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Small heat shock proteins are the well-known regulators of the cytoskeleton integrity, yet their complexes with actin-binding proteins are underexplored. Filamin C, a dimeric 560 kDa protein, abundant in cardiac and skeletal muscles, crosslinks actin filaments and contributes to Z-disc formation and membrane-cytoskeleton attachment. Here, we analyzed the interaction of a human filamin C fragment containing immunoglobulin-like domains 22–24 (FLNC 22-24) with five small heat shock proteins (HspB1, HspB5, HspB6, HspB7, HspB8) and their α-crystallin domains. On size-exclusion chromatography, only HspB7 or its α-crystallin domain formed complexes with FLNC 22-24. Despite similar isoelectric points of the small heat shock proteins analyzed, only HspB7 and its α-crystallin domain interacted with FLNC 22-24 on native gel electrophoresis. Crosslinking with glutaraldehyde confirmed the formation of complexes between HspB7 (or its α-crystallin domain) and the filamin С fragment, inhibiting intersubunit FLNC crosslinking. These data are consistent with the structure modeling using Alphafold. Thus, the C-terminal fragment (immunoglobulin-like domains 22–24) of filamin C contains the site for HspB7 (or its α-crystallin domain) interaction, which competes with FLNC 22-24 dimerization and its probable interaction with different target proteins. [Display omitted] • Human filamin C fragment containing Ig-like domains 22–24 (FLNC 22-24) was obtained. • Interaction of FLNC 22-24 with five human small heat shock proteins was analyzed. • Among five small heat shock proteins tested, only HspB7 interacted with FLNC. • α-crystallin domain of HspB7 can also interact with FLNC 22-24. • HspB7 inhibited FLNC 22-24 dimerization likely by competing for its dimer interface. [ABSTRACT FROM AUTHOR]

It is postulated that the small heat shock proteins directly interact with actin, affect formation and stabilize actin filaments. To verify this suggestion, we have analyzed interaction of recombinant human small heat shock protein HspB7 with skeletal muscle actin. In blot overlay HspB7 binds both G- and F-actin. The sites of interaction are located in the C-terminal large core domain of actin. In the course of ultracentrifugation F-actin and F-actin/tropomyosin complexes were pelleted and trapped HspB7. However, HspB7 pelleting was nonspecific and saturation was not achieved even at very high HspB7 concentration. HspB7 was unable to retard or prevent heat-induced F-actin aggregation. Native gel electrophoresis and chemical crosslinking failed to detect interaction of G-actin with HspB7, although both these methods clearly demonstrated formation of complexes formed by G-actin with DNAse I and cofilin-2. It is concluded that HspB7 is not a genuine actin-binding protein and its effect on actin filaments seems to be determined by interaction of HspB7 with minor regulatory proteins of actin filaments. [Display omitted] • Interaction of HspB7 (cvHsp) with skeletal muscle actin was analyzed. • HspB7 binds both G- and F-actin in immunoblot. • Under ultracentrifugation HspB7 is non-specifically trapped by F-actin. • HspB7 is unable to prevent heat-induced aggregation of F-actin. • In contrast to genuine actin-binding proteins HspB7 does not interact with G-actin. [ABSTRACT FROM AUTHOR]

Ensuring consistent beef eating quality is paramount for meeting consumer demands and sustaining the meat industry. Electrical stimulation (ES) is a post-slaughter intervention used to accelerate post-mortem glycolysis, to avoid cold shortening, to control the tenderization rate of meat through sophisticated physical, chemical and biochemical mechanisms including proteolysis, to improve beef tenderness and to achieve normal pHu that might lead to positive impact on color. This review comprehensively examines the multifaceted effects of ES on beef quality, encompassing factors and settings influencing its efficacy and the underlying biochemical mechanisms revealed using traditional biochemistry methods. It then delves into the molecular pathways modulated by ES, as unveiled by muscle proteomics, aiming to provide a second look and an unprecedented understanding of the underlying biochemical mechanisms through an integrative proteomics analysis of low-voltage ES (LVES) proteomics studies. The proteins changing as a result of ES were gathered in a compendium of 67 proteins, from which 14 were commonly identified across studies. In-depth bioinformatics of this compendium allowed a comprehensive overview of the molecular signatures and interacting biochemical pathways behind electrically stimulated beef muscles. The proteins belong to interconnected molecular pathways including the ATP metabolic process and glycolysis, muscle structure and contraction, heat shock proteins, oxidative stress, proteolysis and apoptosis. Understanding the intricate interplay of molecular pathways behind ES could improve the efficiency of beef production, ensuring consistent meat quality and meeting consumer expectations. The integrative analysis approach performed in this study holds promise for the meat industry's sustainability and competitiveness. • Updated review on the impacts of electrical stimulation (ES) on beef quality determination. • Factors impacting the efficiency of ES have been comprehensively reviewed. • Proteomics allows extending our knowledge on the biochemical mechanisms behind ES beyond traditional biochemistry methods. • First database gathering proteins changing in beef Longissimus muscle as a result of electrical stimulation. • First bioinformatics analysis on electrically stimulated beef muscle proteome revealed interconnected pathways. [ABSTRACT FROM AUTHOR]

Small heat shock protein (HSP20) genes play important roles in biological processes of plants. In this study, a total of 47 CsHSP20 genes, 45 CmHSP20 genes, and 47 ClHSP20 genes were genome-wide identified by ' hmmsearch ' and BLASTP using the latest versions of cucumber, melon, and watermelon genomes, respectively. According to the phylogenetic relationships and predicted subcellular localizations, HSP20s of these three cucurbit species were divided into 8 subfamilies (CI-CIV, CP, ER, M, and PX), in which some HSP20s were closely related with each other based on the collinearity analysis. Specific expression patterns of CsHSP20s were checked in 10 different tissues of cucumber plants. RNA-seq analysis of transcript levels, combined with cis -acting elements and GO enrichment analysis suggested that CsHSP20s were responsive to several different types of abiotic stresses, including chilling, temperature and photoperiod, high temperature and high humidity, and salinity. In conclusion, results of this work not only provided valuable information for exploring the regulating mechanisms of CsHSP20s in responding to abiotic stresses in cucumber, but also shed light on the potentially evolutional relations among cucumber, melon, and watermelon from a perspective of comparative genomics that specified on HSP20 gene families. [ABSTRACT FROM AUTHOR]

Small heat shock protein (sHSP) is a superfamily of molecular chaperone and is found from archaea to human. Recent researches have demonstrated that sHSPs participate in a series of biological processes and are even closely associated with serious diseases. Since sHSP is a very large superfamily and members from different superfamilies exhibit distinct functions, accurate classification of the subfamily of sHSP will be helpful for unrevealing its functions. In the present work, a support vector machine-based method was proposed to classify the subfamily of sHSPs. In the 10-fold cross validation test, an overall accuracy of 93.25% was obtained for classifying the subfamily of sHSPs. The superiority of the proposed method was also demonstrated by comparing it with the other methods. It is anticipated that the proposed method will become a useful tool for classifying the subfamily of sHSPs. • A benchmark dataset was constructed for classifying the superfamily of small heat shock proteins. • The amino acid composition bias was analyzed in different superfamily of small heat shock proteins. • A computational method for classifying the superfamily of small heat shock proteins was proposed. [ABSTRACT FROM AUTHOR]

Physico-chemical properties of HspB6 S10F and P20L mutants with abrogated cardioprotective activity and associated with different forms of cardiomyopathy were analyzed. Under normal conditions both the wild-type HspB6 and its mutants formed small size oligomers (dimers) with apparent molecular weight of 50–60 kDa. Under crowding conditions (0.5 M trimethylamine N-oxide, TMAO) the wild-type HspB6 remained predominantly dimeric or formed small molecular weight complexes, whereas both mutants tended to form high molecular weight complexes. Catalytic subunit of cAMP-dependent protein kinase phosphorylated the wild-type HspB6 and its S10F mutant with comparable rate. The rate of P20L mutant phosphorylation was higher than that of the wild-type HspB6. S10F and P20L mutations did not affect interaction of phosphorylated HspB6 with universal adapter proteins 14-3-3. The wild-type HspB6 was resistant to heat-induced denaturation and aggregation, whereas both its mutants were denatured and started to aggregate at temperature much lower than its wild-type counterpart. Titration with fluorescent probe bis-ANS was accompanied by larger increase of fluorescence in the case of both mutants than in the case of the wild-type HspB6. Both mutants possessed higher chaperone-like activity than the wild-type protein. It is concluded that both S10F and P20L mutations are accompanied by increase of hydrophobicity of the very N-terminal region of HspB6 leading to increased aggregation at elevated temperature, formation of large complexes under crowding conditions and increased chaperone-like activity measured in vitro. Increased hydrophobicity and self-association can affect substrate specificity and interaction with certain target proteins thus leading to decrease or complete abrogation of cardioprotective activity. Image 1 • S10F and P20L mutants abrogating cardioprotection of HspB6 were investigated. • Under crowding conditions mutations promote formation of large HspB6 oligomers. • P20L mutation increases the rate of HspB6 phosphorylation by protein kinase A. • Both mutations increase chaperone-like activity of HspB6 in vitro • Mutations increase N-terminal domain hydrophobicity thus affecting HspB6 properties. [ABSTRACT FROM AUTHOR]

• HCV NS3 has antigenic properties and plays a major role in viral clearance. • Small Hsp27 acts as a protein chaperone, an antioxidant and an immunostimulator. • Histidine-rich R9 and Cady-2 CPPs are able to deliver DNA and proteins, respectively. • Combination of Hsp27, HR9 and Cady-2 significantly improve HCV NS3-specific immunity. To improve an effective hepatitis C virus (HCV) therapeutic vaccine, induction of a strong and long term HCV antigen-specific immune response is an important parameter. HCV non-structural protein 3 (NS3) has antigenic properties and plays a major role in viral clearance. In this study, DNA constructs encoding HCV NS3 and heat shock protein 27 (Hsp27)-NS3 genes, and the recombinant (r) NS3 and rHsp27-NS3 proteins complexed with HR9 and Cady-2 cell penetrating peptides (CPPs) were utilized to evaluate antibody, cytokine and Granzyme B secretion in mice. Herein, the formation of NS3 and Hsp27-NS3 DNA/ HR9 CPP complexes were revealed by gel retardation assay and protection against DNase and protease. Cady-2 peptide was used to form the nanoparticles with rNS3 and rHsp27-NS3 proteins. The size and charge of the nanoparticles were confirmed by SEM and Zetasizer instruments. Next, in vitro transfection of the nanoparticles was assessed by flow cytometry and western blotting. Finally, humoral and cellular immune responses were evaluated using different modalities in mice. Our data showed that HR9 and Cady-2 could form stable nanoparticles with DNA and proteins, respectively and enhance their delivery into HEK-293 T cells in a non-covalent approach. Furthermore, the heterologous Hsp27-NS3 DNA + HR9 prime/rHsp27-NS3+Cady-2 protein boost elicited a higher Th1 cellular immune response with a predominant IgG2a, IgG2b, IFN-γ profile and strong Granzyme B secretion than those induced by other groups. Briefly, the combination of a natural adjuvant (Hsp27) and CPPs (HR9 and Cady-2) could significantly stimulate effective immune responses as a promising approach for development of HCV therapeutic vaccines. [ABSTRACT FROM AUTHOR]

Abstract Utility of Mycobacterium indicus pranii (MIP) as a multistage vaccine against mycobacterial infections demands identification of its protective antigens. We explored antigenicity and immunogenicity of a candidate protein MIP _05962 that depicts homology to HSP18 of M. leprae and antigen1 of Mycobacterium tuberculosis. This protein elicited substantial antibody response in immunized mice along with modulation of cellular immune response towards protective Th1 type. Both CD4+ and CD8+ subsets from immunized mice produced hallmark protective cytokines, IFN-γ, TNF-α and IL-2. This protein also enhanced the CD4+ effector memory that could act as first line of defence during infections. These results point to MIP _05962 as a protective antigen that contributes, in conjunction with others, to the protective immunity of this live vaccine candidate. [ABSTRACT FROM AUTHOR]

Postmortem aging is a value-adding process and has been extensively practiced by the global meat industry for years. The rate and extent of aging impacts on meat quality characteristics are greatly affected by various biochemical/physiological changes occurring during the pre-rigor phase through post-rigor aging processes. This should also mean that the positive aging impacts on eating quality attributes can be further maximized through establishing specific post-harvest aging strategies. In this review, we propose the smart-aging concept, which is to develop innovative template strategies through identifying optimal aging regimes to maximize positive aging impacts on meat quality and value. The concept requires a good understanding of the physical, biochemical and post-harvest factors that affect the aging of beef. This knowledge coupled with the ability to non-invasively determine muscle composition early postmortem will create opportunities to tailor the process of muscle conversion to meat and the subsequent aging processes to deliver meat with consistent and improved eating qualities and functionality. [ABSTRACT FROM AUTHOR]

Chaperones belonging to the small heat shock protein (sHSP) family are ubiquitous and exhibit elevated expression under stresses conditions to protect proteins against aggregation, thereby contributing to the stress tolerance of the organism. Tropical plants are constantly exposed to high temperatures, and the mechanisms by which these plants tolerate heat stress are of foremost importance to basic science as well as applied agrobiotechnology. Therefore, this study aims to characterize sHSPs from different organelles from sugarcane, an important crop that is associated with sugar and bioenergy production. An expression sequence tag database of sugarcane was searched, and sHsp genes of mitochondrial and chloroplast organelles were selected and cloned. The proteins were expressed in Escherichia coli and isolated and purified by two chromatographic steps with high purity as single species. Circular dichroism and fluorescence spectroscopy showed that both proteins were purified in their folded states with a predominant β-sheet secondary structure. Determination of the molecular weight, diffusion coefficient and Stokes radius parameters showed that both chaperones form large spherical-like oligomers in solution. The two sHSPs had different oligomeric states and substrate specificities. The mitochondrial sHSP was a 20-mer with ability to protect model substrates that differ from that of the 16-meric sHSP from chloroplasts. These results indicate that both sHSPs are key agents to protect against stress confirming the importance of the great diversity of sHSP chaperones in plants for homeostasis maintenance. Moreover, to our knowledge, this is the first report about small HSPs from sugarcane organelles. [ABSTRACT FROM AUTHOR]

Physico-chemical properties of G154S, R157H and A171T mutants of αB-crystallin (HspB5) associated with congenital human diseases including certain myopathies and cataract were investigated. Oligomers formed by G154S and A171T mutants have the size and apparent molecular weight indistinguishable from those of the wild-type HspB5, whereas the size of oligomers formed by R157H mutant is slightly smaller. All mutants are less thermostable and start to aggregate at a lower temperature than the wild-type protein. All mutants effectively interact with a triple phosphomimicking mutant of HspB1 and form large heterooligomeric complexes of similar composition. All mutants interact with HspB6 forming heterooligomeric complexes with size and composition dependent on the molar ratio of two proteins. The wild-type HspB5 and its G154S and A171T mutants form only high molecular weight (300–450 kDa) heterooligomeric complexes with HspB6, whereas the R157H mutant forms both high and low (∼120 kDa) molecular weight complexes. The wild-type HspB5 and its G154S and A171T mutants form two types of heterooligomers with HspB4, whereas R157H mutant effectively forms only one type of heterooligomers with HspB4. G154S and A171T mutants have lower chaperone-like activity than the wild-type protein when subfragment S1 of myosin or β L -crystallin are used as a model substrates. With these substrates, the R157H mutant shows equal or higher chaperone activity than the wild-type HspB5. We hypothesize that the mutations in the C-terminal region modulate the binding of the IP(I/V) motif to the core α-crystallin domain. The R157H mutation is located in the immediate proximity of this motif. Such modulation could cause altered interaction of HspB5 with partners and substrates and eventually lead to pathological processes. [ABSTRACT FROM AUTHOR]

It is becoming evident that small heat shock proteins (sHsps) are important players of protein homeostasis system. Their ability to bind misfolded proteins may play a crucial role in preventing protein aggregation in cells. The remarkable structural plasticity of sHsps is considered to underlie the mechanism of their activity. However, all our knowledge of the anti-aggregation functioning of sHsps is based on data obtained in vitro in media greatly different from the cellular highly crowded milieu. The present review highlights available data on the effect of crowding on the anti-aggregation activity of sHsps. There is some evidence that crowding affects conformation and dynamics of sHsps oligomers as well as their anti-aggregation properties. Crowding stimulates association of sHsp—client protein complexes into large-sized aggregates thus diminishing the apparent anti-aggregation activity of sHsps. Nevertheless, it is also shown that complexes between suboligomers (dissociated forms) of sHsps and client proteins may be stabilized and exist for longer period of time under crowded conditions. Moreover, crowding may retard the initial stages of aggregation which correspond to the formation of sHsp-containing nuclei and their clusters. Thus, dissociation of sHsps into suboligomers appears to be an important feature for the anti-aggregation activity of sHsps in crowded media. [ABSTRACT FROM AUTHOR]

Summary By interacting with hundreds of protein partners, 14-3-3 proteins coordinate vital cellular processes. Phosphorylation of the small heat shock protein, HSPB6, within its intrinsically disordered N-terminal domain activates its interaction with 14-3-3, ultimately triggering smooth muscle relaxation. After analyzing the binding of an HSPB6-derived phosphopeptide to 14-3-3 using isothermal calorimetry and X-ray crystallography, we have determined the crystal structure of the complete assembly consisting of the 14-3-3 dimer and full-length HSPB6 dimer and further characterized this complex in solution using fluorescence spectroscopy, small-angle X-ray scattering, and limited proteolysis. We show that selected intrinsically disordered regions of HSPB6 are transformed into well-defined conformations upon the interaction, whereby an unexpectedly asymmetric structure is formed. This structure provides the first atomic resolution snapshot of a human small HSP in functional state, explains how 14-3-3 proteins sequester their regulatory partners, and can inform the design of small-molecule interaction modifiers to be used as myorelaxants. [ABSTRACT FROM AUTHOR]

Neurodegenerative diseases (NDs) are aging-related diseases that involve the death of neurons in the brain. Dysregulation of protein homeostasis leads to the production of toxic proteins or the formation of aggregates, which is the pathological basis of NDs. Small heat shock proteins (HSPB) is involved in the establishment of a protein quality control (PQC) system to maintain cellular homeostasis. HSPB can be secreted into the extracellular space and delivered by various routes, especially extracellular vehicles (EVs). HSPB plays an important role in influencing the aggregation phase of toxic proteins involved in heat shock transcription factor (HSF) regulation, oxidative stress, autophagy and apoptosis pathways. HSPB conferred neuroprotective effects by resisting toxic protein aggregation, reducing autophagy and reducing neuronal apoptosis. The HSPB treatment strategies, including targeted PQC system therapy and delivery of EVs-HSPB, can improve disease manifestations for NDs. This review aims to provide a comprehensive insight into the impact of HSPB in NDs and the feasibility of new technology to enhance HSPB expression and EVs-HSPB delivery for neurodegenerative disease. • Small heat shock proteins can be secreted extracellularly and regulate protein aggregates in neurodegenerative diseases. • HSPB provide neuroprotection by decreasing toxic aggregates, enhancing autophagy and reducing apoptosis. • Targeted HSPB therapeutic strategies can improve symptoms of NDs. • Reducing aggregates could develop new strategies to treat NDs including HSPB-targeted gene therapy, EVs-HSPB nanocarrier delivery, and targeted chimeras. [ABSTRACT FROM AUTHOR]

This study aimed to determine the effect of electrical stimulation and ultimate pH (pH u ) on shear force, myofibrillar protein degradation and small heat shock protein (sHSP) concentrations in M. longissimus lumborum (LL). The LL from both sides of carcasses (n = 15) was excised with low voltage electrical stimulation (ES) applied to an LL muscle from one side, while the opposing LL muscle was not stimulated (NS). Muscles were categorised into low (pH u < 5.8), intermediate (5.8 ≤ pH u < 6.2) and high pH u (pH u ≥ 6.2) and aged for up to 28 days post mortem at − 1.5 °C. High pH u meat tenderised faster which corresponded with the faster degradation of titin and desmin in this group compared with low and intermediate pH u meat. Electrical stimulation significantly affected the variable levels of αβ-crystallin and HSP20 with higher concentrations of these sHSP in ES muscles at later ageing timepoints compared with NS muscles. [ABSTRACT FROM AUTHOR]

Incomplete Kawasaki Disease is a complex disease that often occurs in infants and has substantial coronary artery damage. Its pathogenesis is unclear and lacks specific diagnostic markers. The purpose of our study is to research the mechanism of incomplete Kawasaki Disease use of bioinformatic methods and identify potential biomarkers. We performed weighted gene co‐expression network analysis to analyze the data set GSE68004 and identified modules and genes which were correlated with the disease. Through functional annotation and enrichment analysis, we determined the biological function and signal pathway of these genes. We further used lasso regression and ROC curve to screen genes and determined that the final candidate gene was HSPB11and hsa-miR-155-5p that regulates its expression. Finally, we validated the screened gene using an independent dataset and construct a TF-miRNA network. Through the relationships of TFs and hsa-miR-155-5p, we found is hsa-miR-155-5p closely related to hypoxia-related transcription factors, which may be a new direction in the research of Kawasaki disease. [ABSTRACT FROM AUTHOR]

Small heat shock proteins (sHsps) are conserved across species and are important in stress tolerance. Many sHsps exhibit chaperone-like activity in preventing aggregation of target proteins, keeping them in a folding–competent state and refolding them by themselves or in concert with other ATP-dependent chaperones. Mutations in human sHsps result in myopathies, neuropathies and cataract. Their expression is modulated in diseases such as Alzheimer’s, Parkinson’s and cancer. Their ability to bind Cu 2 + , and suppress generation of reactive oxygen species (ROS) may have implications in Cu 2 + -homeostasis and neurodegenerative diseases. Circulating αB-crystallin and Hsp27 in the plasma may exhibit immunomodulatory and anti-inflammatory functions. αB-crystallin and Hsp20 exhitbit anti-platelet aggregation: these beneficial effects indicate their use as potential therapeutic agents. sHsps have roles in differentiation, proteasomal degradation, autophagy and development. sHsps exhibit a robust anti-apoptotic property, involving several stages of mitochondrial-mediated, extrinsic apoptotic as well as pro-survival pathways. Dynamic N- and C-termini and oligomeric assemblies of αB-crystallin and Hsp27 are important factors for their functions. We propose a “dynamic partitioning hypothesis” for the promiscuous interactions and pleotropic functions exhibited by sHsps. Stress tolerance and anti-apoptotic properties of sHsps have both beneficial and deleterious consequences in human health and diseases. Conditional and targeted modulation of their expression and/or activity could be used as strategies in treating several human disorders. The review attempts to provide a critical overview of sHsps and their divergent roles in cellular processes particularly in the context of human health and disease. [ABSTRACT FROM AUTHOR]

Effect of trimethylamine N-oxide (TMAO), well-known osmolyte, widely used to imitate crowded intracellular conditions, on the quaternary structure of recombinant human small heat shock protein HspB6 (Hsp20) was analyzed by means of size-exclusion chromatography, chemical crosslinking and analytical ultracentrifugation. Consistent with previous reports, in the absence of TMAO unphosphorylated, pseudophosphorylated (S16D mutant) and phosphorylated HspB6 form only small oligomers (presumably dimers). Addition of TMAO to unphosphorylated HspB6 leads to formation of different large oligomers being in equilibrium with dimers. Pseudophosphorylation (S16D mutation) or phosphorylation partially or completely prevent TMAO-induced oligomerization of HspB6. Pseudophosphorylation affects bis-ANS binding suggesting decreased hydrophobicity of HspB6. According to size-exclusion chromatography, TMAO-induced changes of HspB6 oligomerization result in its altered interaction with HspB1 and this effect can be reversed by HspB6 phosphorylation. It is concluded that under conditions of molecular crowding, characteristic for intracellular environment, HspB6 undergoes reversible changes of its oligomeric state which can affect its physiologically important properties and can be delicately regulated by phosphorylation. [ABSTRACT FROM AUTHOR]

Abstract: This study aimed to determine how small heat shock proteins (sHSPs) protect myofibrillar proteins from μ-calpain degradation during ageing. Immunoprecipitation experiments with M. longissimus dorsi (LD) from Angus heifers (n=14) examined the interaction between αβ-crystallin, desmin, titin, HSP20, HSP27 and μ-calpain. Results showed that αβ-crystallin associated with desmin, titin, HSP20, HSP27 and μ-calpain. Exogenous αβ-crystallin reduced desmin and titin degradations in myofibrillar extracts and attenuated μ-calpain activity. In a second experiment, bull LD (n=94) were aged at −1.5°C for up to 28days post mortem. μ-Calpain autolysed faster in high ultimate pH (pHu) meat (pHu ≥6.2) and this was concomitant with the more rapid degradation of titin and filamin in this pHu group. Desmin stability in intermediate pHu meat (pHu 5.8 to 6.19) may be due to the protection of myofibril-bound sHSPs combined with the competitive inhibition of μ-calpain by sHSPs. [Copyright &y& Elsevier]

Abstract: The eating quality of meat is a result of complex interactions between the biological traits and biochemical processes during the conversion of muscle to meat. It was hypothesised that muscles inevitably engage towards apoptotic cell death due to the termination of oxygen and nutrient supply to the muscle following exsanguination. Thus, factors that regulate the process of apoptotic cell death of muscle cells are believed to ultimately influence meat quality. Proteomic studies have associated the regulation of small heat shock proteins (sHSPs) with various meat quality attributes including tenderness, colour, juiciness and flavour. Due to the anti-apoptotic and chaperone functions of sHSPs, they are proposed to be involved with the eating quality of meat. In this review, we discuss the possible chaperone and anti-apoptotic role of sHSPs during the conversion of muscle to meat and consider the repercussions of this on the development of meat tenderness. [Copyright &y& Elsevier]

Abstract: Bull M. longissimus dorsi (n=94) categorised into high (n=28), intermediate (n=14) and low (n=52) ultimate pH (pHu) were aged at −1.5°C for 28days. Shear force was higher and more variable (p<0.05) in intermediate pHu samples during ageing. Titin, filamin and desmin degradation was also less extensive in intermediate pHu samples compared to the other two pH categories. The extent of the decline of HSP20, HSP27 and αβ-crystallin concentrations during post mortem ageing was pHu related such that high pHu meat maintained the highest concentration of small heat shock proteins followed by intermediate and low pHu meat. μ-Calpain autolysis was slowest in intermediate pHu and cathepsin B activities remained consistently low during ageing in this group (p<0.05). Meat toughness in the intermediate pHu group may be attributed to the combination of a larger pool of sHSP with a sub-optimal cathepsin B activity and intermediary μ-calpain activities. [Copyright &y& Elsevier]

Abstract: Some physico-chemical properties of R140G and K141Q mutants of human small heat shock protein HspB1 associated with hereditary peripheral neuropathy were analyzed. Mutation K141Q did not affect intrinsic Trp fluorescence and interaction with hydrophobic probe bis-ANS, whereas mutation R140G decreased both intrinsic fluorescence and fluorescence of bis-ANS bound to HspB1. Both mutations decreased thermal stability of HspB1. Mutation R140G increased, whereas mutation K141Q decreased the rate of trypsinolysis of the central part (residues 5–188) of HspB1. Both the wild type HspB1 and its K141Q mutant formed large oligomers with apparent molecular weight ∼560 kDa. The R140G mutant formed two types of oligomers, i.e. large oligomers tending to aggregate and small oligomers with apparent molecular weight ∼70 kDa. The wild type HspB1 formed mixed homooligomers with R140G mutant with apparent molecular weight ∼610 kDa. The R140G mutant was unable to form high molecular weight heterooligomers with HspB6, whereas the K141Q mutant formed two types of heterooligomers with HspB6. In vitro measured chaperone-like activity of the wild type HspB1 was comparable with that of K141Q mutant and was much higher than that of R140G mutant. Mutations of homologous hot-spot Arg (R140G of HspB1 and R120G of αB-crystallin) induced similar changes in the properties of two small heat shock proteins, whereas mutations of two neighboring residues (R140 and K141) induced different changes in the properties of HspB1. [Copyright &y& Elsevier]

Abstract: Lymphatic filariasis affects approximately 3% of the whole world population. Mass drug administration is currently the major control strategy to eradicate this infection from endemic regions by year 2020. Combination drug treatments are highly efficient in controlling the infection. However, there are no effective vaccines available for human or animal lymphatic filariasis despite the identification of several subunit vaccines. Lymphatic filariasis parasites are multicellular organisms and potentially use multiple mechanisms to survive in the host. Therefore, there is a need to combine two or more vaccine candidate antigens to achieve the desired effect. In this study we combined three well characterized vaccine antigens of Brugia malayi, heat shock protein 12.6 (HSP12.6), Abundant Larval transcript-2 (ALT-2) and tetraspanin large extra cellular loop (TSP-LEL) as a multivalent fusion vaccine. Putative immune individuals carry circulating antibodies against all three antigens. Depletion of these antigen specific antibodies from the sera samples removed the ability of the sera to participate in the killing of B. malayi L3 in an antibody dependent cellular cytotoxicity (ADCC) mechanism. Vaccination trials in mice with a bivalent [HSP12.6+ALT-2 (HA), HSP12.6+TSP-LEL (HT) or TSP-LEL+ALT-2 (TA)] or trivalent [HSP12.6+ALT-2+TSP-LEL (HAT)] vaccines using DNA, protein or heterologous prime boost regimen showed that trivalent HAT vaccine either as protein alone or as heterologous prime boost vaccine could confer significant protection (95%) against B. malayi L3 challenge. Immune correlates of protection suggest a Th1/Th2 bias. These finding suggests that the trivalent HAT fusion protein is a promising prophylactic vaccine against lymphatic filariasis infection in human. [Copyright &y& Elsevier]

Abstract: The members of the small heat shock protein (sHSP) family are molecular chaperones that play major roles in development, stress responses, and diseases, and have been envisioned as targets for therapy, particularly in cancer. The molecular mechanisms that regulate their transcription, in normal, stress, or pathological conditions, are characterized by extreme complexity and subtlety. Although historically linked to the heat shock transcription factors (HSFs), the stress-induced or developmental expression of the diverse members, including HSPB1/Hsp27/Hsp25, αA-crystallin/HSPB4, and αB-crystallin/HSPB5, relies on the combinatory effects of many transcription factors. Coupled with remarkably different cis-element architectures in the sHsp regulatory regions, they confer to each member its developmental expression or stress-inducibility. For example, multiple regulatory pathways coordinate the spatio-temporal expression of mouse αA-, αB-crystallin, and Hsp25 genes during lens development, through the action of master genes, like the large Maf family proteins and Pax6, but also HSF4. The inducibility of Hsp27 and αB-crystallin transcription by various stresses is exerted by HSF-dependent mechanisms, by which concomitant induction of Hsp27 and αB-crystallin expression is observed. In contrast, HSF-independent pathways can lead to αB-crystallin expression, but not to Hsp27 induction. Not surprisingly, deregulation of the expression of sHSP is associated with various pathologies, including cancer, neurodegenerative, or cardiac diseases. However, many questions remain to be addressed, and further elucidation of the developmental mechanisms of sHsp gene transcription might help to unravel the tissue- and stage-specific functions of this fascinating class of proteins, which might prove to be crucial for future therapeutic strategies. This article is part of a Directed Issue entitled: Small HSPs in physiology and pathology. [Copyright &y& Elsevier]

Abstract: The stress responses of most bacteria are thought to involve the upregulation of small heat shock proteins. We describe here some of the most pertinent aspects of small heat shock proteins, to highlight their potential for use in various applications. Bacterial species have between one and 13 genes encoding small heat shock proteins, the precise number depending on the species considered. Major efforts have recently been made to characterize the protein protection and membrane stabilization mechanisms involving small heat shock proteins in bacteria. These proteins seem to be involved in the acquisition of cellular heat tolerance. They could therefore potentially be used to maintain cell viability under unfavorable conditions, such as heat shock or chemical treatments. This review highlights the potential roles of applications of small heat shock proteins in stabilizing overproduced heterologous proteins in Escherichia coli, purified bacterial small heat shock proteins in protein biochip technology, proteomic analysis and food technology and the potential impact of these proteins on some diseases. This article is part of a Directed Issue entitled: Small HSPs in physiology and pathology. [Copyright &y& Elsevier]

Abstract: A number of neurological and muscular disorders are characterized by the accumulation of aggregate-prone proteins and are referred to as protein deposit or protein conformation diseases. Besides some sporadic forms, most of them are genetically inherited in an autosomal dominant manner, although recessive forms also exist. Although genetically very heterogeneous, some of these diseases are the result of mutations in some members of the mammalian small heat shock protein family (sHSP/HSPB), which are key players of the protein quality control system and participate, together with other molecular chaperones and co-chaperones, in the maintenance of protein homeostasis. Thus, on one hand upregulation of specific members of the HSPB family can exert protective effects in protein deposit diseases, such as the polyglutamine diseases. On the other hand, mutations in the HSPBs lead to neurological and muscular disorders, which may be due to a loss-of-function in protein quality control and/or to a gain-of-toxic function, resulting from the aggregation-proneness of the mutants. In this review we summarize the current knowledge about some of the best characterized functions of the HSPBs (e.g. role in cytoskeleton stabilization, chaperone function, anti-aggregation and anti-apoptotic activities), also highlighting differences in the properties of the various HSPBs and how these may counteract protein aggregation diseases. We also describe the mutations in the various HSPBs associated with neurological and muscular disorders and we discuss how gain-of-toxic function mechanisms (e.g. due to the mutated HSPB protein instability and aggregation) and/or loss-of-function mechanisms can contribute to HSPB-associated pathologies. This article is part of a Directed Issue entitled: Small HSPs in physiology and pathology. [Copyright &y& Elsevier]

Abstract: The expression of small heat shock proteins is tightly regulated during development in multiple organisms. As housekeeping proteins, small heat shock proteins help protect cells from apoptosis, stabilize the cytoskeleton and contribute to proteostasis. Consistently, depletion of one small heat shock protein is usually not detrimental due to a certain level of redundancy between the functions of each small heat shock protein. However, while their stress-induced expression is regulated by heat shock factors, their constitutive expression is under the control of other specific transcription factors, suggesting the existence of very specialized functions. This review focuses on the expression patterns and functions of small heat shock proteins in various organisms during development. This article is part of a Directed Issue entitled: Small HSPs in physiology and pathology. [Copyright &y& Elsevier]

Abstract: Fluorescent chimeras composed of enhanced cyan (or enhanced yellow) fluorescent proteins (ECFP or EYFP) and one of the four human small heat shock proteins (HspB1, HspB5, HspB6 or HspB8) were expressed in E. coli and purified. Fluorescent chimeras were used for investigation of heterooligomeric complexes formed by different small heat shock proteins (sHsp) and for analysis of their subunit exchange. EYFP-HspB1 and ECFP-HspB6 form heterooligomeric complex with apparent molecular weight of ∼280 kDa containing equimolar quantities of both sHsp. EYFP-HspB5 and ECFP-HspB6 formed heterogeneous oligomeric complexes. Fluorescent proteins inside heterooligomeric complexes formed by HspB1/HspB6 and HspB5/HspB6 chimeras are closely located, making possible effective fluorescence resonance energy transfer (FRET). Neither the wild type HspB8 nor its fluorescent chimeras were able to form stable heterooligomeric complexes with the wild type HspB1 and HspB5. Homo- and hetero-FRET was used for analysis of subunit exchange of small heat shock proteins. The apparent rate constant of subunit exchange was temperature-dependent and was higher for HspB6 forming small oligomers than for HspB1 forming large oligomers. Replacement induced by homologous subunits was more rapid than the replacement induced by heterologous subunits of small heat shock proteins. Fusion of fluorescent proteins might affect oligomeric structure of small heat shock proteins, however fluorescent chimeras can be useful for investigation of heterooligomeric complexes formed by sHsp and for analysis of kinetics of their subunit exchange. [Copyright &y& Elsevier]

Abstract: The ability of photosynthetic organisms to adapt to increases in environmental temperatures is becoming more important with climate change. Heat stress is known to induce heat-shock proteins (HSPs) many of which act as chaperones. Traditionally, it has been thought that protein denaturation acts as a trigger for HSP induction. However, increasing evidence has shown that many stress events cause HSP induction without commensurate protein denaturation. This has led to the membrane sensor hypothesis where the membrane’s physical and structural properties play an initiating role in the heat shock response. In this review, we discuss heat-induced modulation of the membrane’s physical state and changes to these properties which can be brought about by interaction with HSPs. Heat stress also leads to changes in lipid-based signaling cascades and alterations in calcium transport and availability. Such observations emphasize the importance of membranes and their lipids in the heat shock response and provide a new perspective for guiding further studies into the mechanisms that mediate cellular and organismal responses to heat stress. [Copyright &y& Elsevier]

Abstract: Motor neuron diseases (MNDs) are neurodegenerative disorders that specifically affect the survival and function of upper and/or lower motor neurons. Since motor neurons are responsible for the control of voluntary muscular movement, MNDs are characterized by muscle spasticity, weakness and atrophy. Different susceptibility genes associated with an increased risk to develop MNDs have been reported and several mutated genes have been linked to hereditary forms of MNDs. However, most cases of MNDs occur in sporadic forms and very little is known on their causes. Interestingly, several molecular mechanisms seem to participate in the progression of both the inherited and sporadic forms of MNDs. These include cytoskeleton organization, mitochondrial functions, DNA repair and RNA synthesis/processing, vesicle trafficking, endolysosomal trafficking and fusion, as well as protein folding and protein degradation. In particular, accumulation of aggregate-prone proteins is a hallmark of MNDs, suggesting that the protein quality control system (molecular chaperones and the degradative systems: ubiquitin–proteasome-system and autophagy) are saturated or not sufficient to allow the clearance of these altered proteins. In this review we mainly focus on the MNDs associated with disturbances in protein folding and protein degradation and on the potential implication of a specific class of molecular chaperones, the small heat shock proteins (sHSPs/HSPBs), in motor neuron function and survival. How boosting of specific HSPBs may be a potential useful therapeutic approach in MNDs and how mutations in specific HSPBs can directly cause motor neuron degeneration is discussed. [Copyright &y& Elsevier]

Abstract: A small heat shock gene of Lactobacillus plantarum strain WCFS1 was deleted using a Cre-lox based system. Compared to the wild type, the ∆hsp 18.55 mutant strain displayed a similar growth rate when cultivated either under optimal temperature or under different stress conditions such as heat, low pH and salt stress. However, a longer lag phase was observed when the ∆hsp 18.55 mutant strain was cultivated under short intense heat stress (50 °C). This suggests that the hsp 18.55 gene of L. plantarum may be involved in recovery of L. plantarum stressed cells in the early stage of high temperature stress. In addition, morphology of the mutant cells, investigated by scanning electron microscopy, revealed that cells clumped together and had rough surfaces, and that some of the cells had a shrunken empty appearance, which clearly contrasted with the characteristic rod-shaped, smooth-surface morphology of control L. plantarum cells. Furthermore, inactivation of the hsp 18.55 gene affected membrane fluidity and physicochemical surface properties of L. plantarum WCFS1. [Copyright &y& Elsevier]

Abstract: We applied different methods (differential scanning calorimetry, circular dichroism, Fourier transform infrared spectroscopy, and intrinsic fluorescence) to investigate the thermal-induced changes in the structure of small heat shock protein Hsp22. It has been shown that this protein undergoes thermal-induced unfolding that occurs within a very broad temperature range (from 27 °C to 80 °C and above), and this is accompanied by complete disappearance of α-helices, significant decrease in β-sheets content, and by pronounced changes in the intrinsic fluorescence. The results confirm predictions that Hsp22 belongs to the family of intrinsically disordered proteins (IDP) with certain parts of its molecule (presumably, in the α-crystallin domain) retaining folded structure and undergoing reversible thermal unfolding. The results are also discussed in terms of downhill folding scenario. [Copyright &y& Elsevier]

Abstract: In hereditary cerebral hemorrhage with amyloidosis of the Dutch type (HCHWA-D), severe cerebral amyloid angiopathy (CAA) is associated with an inflammatory reaction. Small heat shock proteins (sHsps) are molecular chaperones and association of HspB8 with CAA in HCHWA-D has been observed. The aims of this study were to investigate (1) if other sHsps are associated with the pathological lesions in HCHWA-D brains, (2) if the amyloid-β protein (Aβ) increases production of sHsps in cultured cerebral cells and (3) if sHsps are involved in the cerebral inflammatory processes in both Alzheimer''s disease (AD) and HCHWA-D. We conclude that Hsp20, HspB8 and HspB2 are present in CAA in HCHWA-D, and that Aβ did not affect cellular sHsps expression in cultured human brain pericytes and astrocytes. In addition, we demonstrated that Hsp20, HspB2 and HspB8 induced interleukin-6 production in cultured pericytes and astrocytes, which could be antagonized by dexamethasone, whereas other sHsps and Aβ were inactive, suggesting that sHsps may be among the key mediators of the local inflammatory response associated with HCHWA-D and AD lesions. [Copyright &y& Elsevier]

[Display omitted] Increasing evidence points towards using extracellular vesicles (EVs) as a therapeutic strategy in neurodegenerative diseases such as multiple sclerosis, Parkinson's, and Alzheimer's disease. EVs are nanosized carriers that play an essential role in intercellular communication and cellular homeostasis by transporting an active molecular cargo, including a large variety of proteins. Recent publications demonstrate that small heat shock proteins (HSPBs) exhibit a beneficial role in neurodegenerative diseases. Moreover, it is defined that HSPBs target the autophagy and the apoptosis pathway, playing a prominent role in chaperone activity and cell survival. This review elaborates on the therapeutic potential of EVs and HSPBs, in particular HSPB1 and HSPB8, in neurodegenerative diseases. We conclude that EVs and HSPBs positively influence neuroinflammation, central nervous system (CNS) repair, and protein aggregation in CNS disorders. Moreover, we propose the use of HSPB-loaded EVs as advanced nanocarriers for the future development of neurodegenerative disease therapies. [ABSTRACT FROM AUTHOR]

Abstract: We isolated and characterized two members of the α-crystallin/sHsp family, SnoHsp19.5 and SnoHsp20.8 from Sesamia nonagrioides (Lepidoptera: Noctuidae). The cDNAs encoded proteins of 174 and 185 amino acids, with calculated molecular weights of 19.5 and 20.8kDa, respectively. The deduced amino acid sequences of SnoHsp19.5 and SnoHsp20.8 showed highest homology to Hsp19.7 of Mamestra brassicae and to Bombyx mori Hsp20.4, respectively. Expression patterns of SnoHsp19.5 and SnoHsp20.8 in non-diapausing individuals under different environmental conditions (heat or cold) showed different accumulation profiles for the two genes after heat and cold treatment. SnoHsp19.5 was consistently expressed, while SnoHsp20.8 gene was down-regulated in deep diapause and was up-regulated at the termination of diapause. Our results suggest that these two genes play distinctive roles in the regulation of diapause. [Copyright &y& Elsevier]

Abstract: Small heat shock proteins (sHsps) exist in almost all organisms. Most organisms have more than one sHsp, but their number can be as high as 65, as found in the eukaryote, Vitis vinifera. The function of sHsps is well-known; they confer thermotolerance to cellular cultures and proteins in cellular extracts during prolonged incubations at elevated temperatures. This demonstrates the ability of sHsps to protect cellular proteins, and to maintain cellular viability under conditions of intensive stress, such as heat shock or chemical treatments. sHsps have several properties that distinguish them from heat shock proteins (Hsps): they function as ATP-independent chaperones, require the flexible assembly and reassembly of oligomeric complex structures for their activation, and exhibit a wide range of substrate-binding capacities. Recent studies indicate that sHsps have important biological functions in thermostability, disaggregation, and proteolysis inhibition. These functions can be harnessed for various applications, including nanobiotechnology, proteomics, bioproduction, and bioseparation. In this review, we discuss the properties and diversity of microbial sHsps, as well as their potential uses in the biotechnology industry. [Copyright &y& Elsevier]

Abstract: The small heat shock proteins (sHsps) are a family of molecular chaperones defined by an alpha-crystallin domain that is important for sHsps oligomerization and chaperone activity. sHsps perform many physiological functions including the maintenance of the cellular cytoskeleton, the regulation of protein aggregation and modulate cell survival in a number of cell types including glial and neuronal cells. Many of these functions have been implicated in disease processes in the CNS and indeed sHsps are considered targets for disease therapy. Despite this, there is no study that systematically and comparatively characterized sHsps expression in the CNS. In the present study we have analyzed the expression of this gene family in the mouse brain by reverse-transcriptase polymerase chain reaction (RT-PCR), in situ hybridization and Western blotting. Gene expression analysis of the 10 known members of mammalian sHsps confirms the presence of 5 sHsps in the CNS. A distinct white matter specific expression pattern for HspB5 and overlapping expression of HspB1 and HspB8 in the lateral and dorsal ventricles of the brain is observed. We confirm protein expression of HspB1, HspB5, HspB6 and HspB8 in the brain. Further subcellular fractionation of brain and synaptosomes details a distinct subcompartment-specific association and detergent solubility of sHsps. This biochemical signature is indicative of an association with synaptic and other neural specializations. This observation will help one understand the functional role played by sHsps during physiology and pathology in the CNS. [Copyright &y& Elsevier]

Abstract: In addition to heat shock (HS), a proline analog, azetidine-2-carboxylic acid (Aze), also induces heat shock protein (HSP) synthesis. Here, we describe our further characterization of the induced effects by Aze treatment in soybean seedlings. Northern blot analyses revealed transcription of the soybean HS genes hsp70 and hsc70 as well as hspl7.5 activated after 6h of Aze incubation, as reported previously. However, Aze-induced activation of HSP genes was not attenuated after transferring seedlings pretreated with Aze to shaking buffer, whereas the activation was decreased when seedlings were treated with Aze and then proline. Besides triggering an HS-like response, Aze treatment triggered a continuous increase of cellular leakage in soybean seedlings at room temperature. Nonlethal HS pretreatment protected seedlings from cellular leakage induced by Aze and allowed them to survive subsequent lethal HS treatment. As well, 10mM proline prevented electrolyte leakage after 6h of 10mM Aze treatment and increased the survival of seedlings under lethal treatment. These results indicate that Aze treatment causes a permeability change in the plasma membrane by leading to cellular leakage before accumulation of HSPs. In vitro chaperone activity of 10mM Aze-induced sHSP-CIs (class I small HSPs) with the model substrate citrate synthase was less efficient than that from 2h of 40°C treatment. [Copyright &y& Elsevier]

Abstract: Small heat shock proteins (HSPs) have been shown to confer thermotolerance in many organisms. Here, we demonstrate that small HSPs (sHSPs) can also be involved in development of thermotolerance in Pisolithus sp. In heat shock response, Pisolithus isolate RV82 synthesized proteins of molecular mass 28, 26 and 15–18kDa. These group of proteins are synthesized when mycelial mass are exposed to heat shock temperature (42°C) for short period (30min) and incubated back at 28°C, the optimal temperature for growth. Our results show sHSPs are an important biochemical alteration in ectomycorrhizal fungi under thermal stress. [Copyright &y& Elsevier]

Small heat shock proteins (sHSPs) are widely distributed, and their function and diversity of structure have been much studied in the field of molecular chaperones. In plants, which frequently have to cope with hostile environments, sHSPs are much more abundant and diverse than in other forms of life. In response to high temperature stress, sHSPs of more than twenty kinds can make up more than 1% of soluble plant proteins. We isolated a genomic clone, NtHSP18.3, from Nicotiana tabacum that encodes the complete open reading frame of a cytosolic class I small heat shock protein. To investigate the function of NtHSP18.3 in vitro, it was overproduced in Escherichia coli and purified. The purified NtHSP18.3 had typical molecular chaperone activity as it protected citrate synthase and luciferase from high temperature-induced aggregation. When E. coli celluar proteins were incubated with NtHSP18.3, a large proportion of the proteins remained soluble at temperatures as high as 70°C. Native gel analysis suggested that NtHSP18.3 is a dodecameric oligomer as the form present and showing molecular chaperone activity at the condition tested. Binding of bis-ANS to the oligomers of NtHSP18.3 indicated that exposure of their hydrophobic surfaces increased as the temperature was raised. Taken together, our data suggested that NtHSP18.3 is a molecular chaperone that functions as a dodecameric complex and possibly in a temperature-induced manner. [ABSTRACT FROM AUTHOR]

Abstract: Increased expression of small heat shock proteins (sHSPs) is known to be a key regulatory mechanism in extending tolerance to a variety of environmental stresses. In the present study, a full-length cDNA clone encoding a member of the α-crystallin/sHSP family was isolated and characterized from the endoparasitic wasp Venturia canescens (Hymenoptera: Ichneumonidae). Western blot analysis indicated that the mature protein has a mass of about 35 kDa (Vc_sHSP35). Sequence analysis of RT-PCR products revealed that two transcript forms of the gene are expressed in different developmental stages and tissues of the wasp, with the longer form likely to contain an unspliced intron sequence. Furthermore, gene expression was analysed in ovaries of V. canescens wasps from two genetically different lines after exposure to different temperatures (heat or cold shock and heat or cold acclimation, respectively). Wasps from both lines principally showed the same cold induced change in expression of the shorter transcript form (Vc_sHSP35-2). However, expression levels were higher in wasps from one line compared to the other. These results are discussed in relation to the environmental stress resistance and molecular ecology of both V. canescens lines. [Copyright &y& Elsevier]

Abstract: The objective of the present study was to evaluate the relationship between small heat shock proteins (sHSP) degradation and tenderness development of beef loins at different ultimate pH (pHu). A total of twelve loins (M. longissimus dorsi) from steers were obtained at 1day post mortem. Shear force and proteolysis of each loin were analyzed at 1 and 28days post mortem. The loins at intermediate pHu (5.8 to 6.0) showed more variation in tenderness compared to the loins at low pHu (<5.8), where few samples were still tough (>10kgF) at 28days. The intact sHSP20 was more pronounced (P <0.05) in the intermediate pHu loin compared to the low pHu counterpart. Further, high correlations between the degradation of both sHSP and myofibrillar proteins were observed (e.g. r =0.94; degraded sHSP27 and degraded desmin). The result of this study suggests that the extent of sHSP degradation could be an indicator of myofibrillar protein degradation and tenderness. [Copyright &y& Elsevier]

Abstract: The recently published paper by Wettstein et al. (2012) reviews the data of literature dealing with participation of small heat shock proteins (sHsp) in cytoskeleton regulation. Analyzing the effect of sHsp on microfilaments, the authors come to conclusion that depending on phosphorylation HspB1 can function as barbed-end-capping protein and can prevent aggregation of F-actin under stress conditions. The modern data do not confirm all these suggestions. We propose that stabilization effect of HspB1 on microfilaments is due to HspB1 interaction with partially unfolded actin or with genuine actin-binding proteins. In addition, HspB1 can exert its stabilizing effect on F-actin by modulating other elements of the cytoskeleton (intermediate filaments and microtubules) or by controlling homeostasis (for instance, redox state). Without being genuine actin-binding proteins, HspB1 and HspB6 predominantly protect microfilaments via an indirect mechanism that is yet to be characterized. [Copyright &y& Elsevier]

The objective of this study was to determine the proteolytic changes of myofibrillar and small heat shock proteins (HSPs) in different muscles during aging and to evaluate their relevance to meat quality attributes. From 8 beef carcasses, longissimus lumborum (LL), semimembranosus (SM), and psoas major (PM) muscles were obtained, cut into sections and assigned to various aging periods up to 23d. PM exhibited limited aging potential in quality developments shown by lower extents of shear force, water-holding capacity (WHC), and proteolytic changes, including calpain 1 autolysis, troponin T, and HSP27 compared to LL and SM. Conversely, LL had an increase in tenderization and WHC, which was accompanied with more extended calpain 1 autolysis, proteolysis and HSP27 degradation, compared with other muscles. The results of this study suggest that postmortem proteolytic changes of myofibrillar proteins, small HSPs and apoptotic factors occur in a muscle-specific manner, which is likely attributed to different rate and extent of meat quality developments of each muscle during aging. • LL showed extended proteolysis and μ-calpain autolysis during aging. • LL showed an increase in HSP27 degradation and early caspase 3 cleavage. • PM had less extent of proteolysis in troponin T and HSP27 compared to LL. • Apoptotic changes and subsequent meat tenderization occur in a muscle-specific manner. [ABSTRACT FROM AUTHOR]

Heat shock protein 27 regulates homeostasis of skeletal and cardiac muscle proteins in various stressful states including diabetes and exercise. Aerobic exercise can inhibit or ameliorate cardiac structural abnormality and dysfunction in diabetic cardiomyopathy. The aim of this study was to evaluate the role of HSP27 in aerobic exercise improving cardiac diastolic dysfunction in type 2 diabetic rats. Forty male Sprague-Dawley rats were randomly divided into the following groups: control, control + aerobic exercise, diabetic, and diabetic + aerobic exercise. Diabetes was induced by feeding with a high-fat high-sugar diet for 7-weeks followed by a single intraperitoneal injection of streptozotocin (30 mg/kg) in male rats. Moderate aerobic exercise training was performed on a treadmill for 8 weeks after induction of diabetes. Aerobic exercise increased left ventricular end-diastolic internal diameter, left ventricular end-diastolic volume, myocardial HSP27 protein expression, HSP27-S82 phosphorylation levels, pHSP27-titin binding and improved cardiac muscle fibre alignment in diabetic rats. Our study indicates that moderate aerobic exercise increases HSP27 activation, improves cardiomyocyte fibre alignment and restores cardiac diastolic function. [ABSTRACT FROM AUTHOR]

• Identification and in silico characterization of choloroplast specific HSP26 allelic variants in wild relatives of wheat. • A novel HSP26 gene discovered in DD-genome assembly of diploid progenitor wheat, the orthologue of which -is not present in hexaploid wheat genome. • First report -on the isolation of a novel chloroplast specific sHSP26 protein from Ae. tauschii accession pau14376 (GenBank:MN119514). • A 9-bp indel in TmHSP26-1(GA) translated into a deletion of SPM amino acid segment in chloroplast specific conserved consensus region III. • High degree of divergence in nucleotide sequence between cultivated and wild species appeared in the form of higher ω values (Ka/Ks >1). Heat shock proteins are an important class of molecular chaperones known to impart tolerance under high temperature stress. sHSP26, a member of small heat shock protein subfamily is specifically involved in protecting plant's photosynthetic machinery. The present study aimed at identifying and characterizing sequence and structural variations in sHSP26 from genetically diverse progenitor and non-progenitor species of wheat. In silico analysis identified three paralogous copies of TaHSP26 to reside on short arm of chromosome 4A while one homeologue each was localized on long arm of chromosome 4B and 4D of cultivated bread wheat. Wild DD-genome donor Aegilops tauschii carried an additional sHSP26 gene (AET4Gv20569400) which was absent in the cultivated DD genome of bread wheat. In vitro amplification of this novel gene in wild accessions of Ae. tauschii and synthetic hexaploid wheat but not in cultivated bread wheat validated this finding. Further, significant length polymorphism could be identified in exon1 from diverse sHSP26 sequences. Multiple sequence alignment of procured sequences revealed numerous sSNPs and nsSNPs. D3A, P125 L, Q242 K were designated as homeolog specific- while A49 G as non-progenitor specific amino acid replacements. A 9-bp indel in TmHSP26-1(GA) translated into a deletion of SPM amino acid segment in chloroplast specific conserved consensus region III. High degree of divergence in nucleotide sequence between cultivated and wild species appeared in the form of higher ω values (Ka/Ks >1) indicating positive selection during the course of evolution. Phylogenetic analysis elucidated ancestral relationships between wheat sHSP26 proteins and orthologous proteins across plant kingdom. Overall, data mining approach may be employed as an effective pre-breeding strategy to identify and mobilize novel stress responsive genes and distinct allelic variants from wider germplasm collections of wheat to enhance climate resilience of present day elite wheat cultivars. [ABSTRACT FROM AUTHOR]