19 results on '"Hedtke T"'
Search Results
2. Unique molecular networks: Formation and role of elastin cross-links
- Author
-
Schmelzer, C.E.H., Hedtke, T., Heinz, A., and Publica
- Abstract
Elastic fibers are essential assemblies of vertebrates and confer elasticity and resilience to various organs including blood vessels, lungs, skin, and ligaments. Mature fibers, which comprise a dense and insoluble elastin core and a microfibrillar mantle, are extremely resistant toward intrinsic and extrinsic influences and maintain elastic function over the human lifespan in healthy conditions. The oxidative deamination of peptidyl lysine to peptidyl allysine in elastin's precursor tropoelastin is a crucial posttranslational step in their formation. The modification is catalyzed by members of the family of lysyl oxidases and the starting point for subsequent manifold condensation reactions that eventually lead to the highly cross-linked elastomer. This review summarizes the current understanding of the formation of cross-links within and between the monomer molecules, the molecular sites, and cross-link types involved and the pathological consequences of abnormalities in the cross-linking process.
- Published
- 2020
3. Direct three-dimensional imaging for morphological analysis of electrospun fibers with laboratory-based Zernike X-ray phase-contrast computed tomography
- Author
-
Santos de Oliveira, C., Gonzalez, A.T., Hedtke, T., Kürbitz, T., Heilmann, A., Schmelzer, C.E.H., Martins de S. e Silva, J., and Publica
- Abstract
Electrospinning is a well-established and widely used method for the production of protein-based fibrous biomaterials. The visualization of the morphology and the characterization of sample features related to the three-dimensional (3D) structure, like the porosity and fibers thickness, is crucial for the design and fabrication of tailor-made and application-optimized materials. Here, we evaluated the benefits of using 3D X-ray imaging in a laboratory setup with a resolution in the sub-micrometer range for the characterization of electrospun gelatin fibrous mats. We used phase-contrast X-ray computed tomography at the nanoscale (nano-CT) for the evaluation of the time-course morphological changes of the mats induced by chemical cross-linking of the gelatin fibers. We present an image processing protocol that enables the segmentation of the fibers and quantification of the mats porosity, the analysis of the shape and size of the pores, and of the fibers thickness and orientation. We compared the results obtained from the processed nano-CT data with those obtained with the conventional methods used for the characterization of electrospun fibrous materials, and we discuss the advantages and limitations of each method when applied to gelatin electrospun samples. Our results reveal that the use of phase-contrast nano-CT provides quick additional and relevant information for the characterization of fibrous mats and, thus, provides beneficial insights for the design and fabrication of novel fibrous materials.
- Published
- 2020
4. Lysyl oxidase-like 2 (LOXL2)-mediated cross-linking of tropoelastin
- Author
-
Schmelzer, C.E.H., Heinz, A., Troilo, H., Lockhart-Cairns, M.P., Jowitt, T.A., Marchand, M.F., Bidault, L., Bignon, M., Hedtke, T., Barret, A., McConnell, J.C., Sherratt, M.J., Germain, S., Hulmes, D.J.S., Baldock, C., Muller, L., and Publica
- Abstract
Lysyl oxidases (LOXs) play a central role in extracellular matrix remodeling during development and tumor growth and fibrosis through cross-linking of collagens and elastin. We have limited knowledge of the structure and substrate specificity of these secreted enzymes. LOXs share a conserved C-terminal catalytic domain but differ in their N-terminal region, which is composed of 4 repeats of scavenger receptor cysteine-rich (SRCR) domains in LOX-like (LOXL) 2. We investigated by X-ray scattering and electron microscopy the low-resolution structure of the full-length enzyme and the structure of a shorter form lacking the catalytic domain. Our data demonstrate that LOXL2 has a rod-like structure with a stalk composed of the SRCR domains and the catalytic domain at its tip. We detected direct interaction between LOXL2 and tropoelastin (TE) and also LOXL2-mediated deamination of TE. Using proteomics, we identified several allysines together with cross-linked TE peptides. The elastin-like material generated was resistant to trypsin proteolysis and displayed mechanical properties similar to mature elastin. Finally, we detected the codistribution of LOXL2 and elastin in the vascular wall. Altogether, these data suggest that LOXL2 could participate in elastogenesis in vivo and could be used as a means of cross-linking TE in vitro for biomimetic and cell-compatible tissue engineering purposes.
- Published
- 2019
5. A comprehensive map of human elastin cross-linking during elastogenesis
- Author
-
Hedtke, T., Schräder, C.U., Heinz, A., Hoehenwarter, W., Brinckmann, J., Groth, T., Schmelzer, C.E.H., and Publica
- Abstract
Elastin is an essential structural protein in the extracellular matrix of vertebrates. It is the core component of elastic fibers, which enable connective tissues such as those of the skin, lungs or blood vessels to stretch and recoil. This function is provided by elastin's exceptional properties, which mainly derive from a unique covalent cross-linking between hydrophilic lysine-rich motifs of units of the monomeric precursor tropoelastin. To date, elastin's cross-linking is poorly investigated. Here, we purified elastin from human tissue and cleaved it into soluble peptides using proteases with different specificities. We then analyzed elastin's molecular structure by identifying unmodified residues, post-translational modifications and cross-linked peptides by high-resolution mass spectrometry and amino acid analysis. The data revealed the presence of multiple isoforms in parallel and a complex and heterogeneous molecular interconnection. We discovered that the same lysine residues in different monomers were simultaneously involved in various cross-link types or remained unmodified. Furthermore, both types of cross-linking domains, Lys-Pro and Lys-Ala domains, participate not only in bifunctional inter- but also in intra-domain cross-links. We elucidated the sequences of several desmosine-containing peptides and the contribution of distinct domains such as 6, 14 and 25. In contrast to earlier assumptions proposing that desmosine cross-links are formed solely between two domains, we elucidated the structure of a peptide that proves a desmosine formation with participation of three Lys-Ala domains. In summary, these results provide new and detailed insights into the cross-linking process, which takes place within and between human tropoelastin units in a stochastic manner.
- Published
- 2019
6. Underpotential Deposition of 3D Transition Metals: Versatile Electrosynthesis of Single-Atom Catalysts on Oxidized Carbon Supports.
- Author
-
Meese AF, Napier C, Kim DJ, Rigby K, Hedtke T, Leshchev D, Stavitski E, Parent LR, and Kim JH
- Abstract
Use of single-atom catalysts (SACs) has become a popular strategy for tuning activity and selectivity toward specific pathways. However, conventional SAC synthesis methods require high temperatures and pressures, complicated procedures, and expensive equipment. Recently, underpotential deposition (UPD) has been investigated as a promising alternative, yielding high-loading SAC electrodes under ambient conditions and within minutes. Yet only few studies have employed UPD to synthesize SACs, and all have been limited to UPD of Cu. In this work, a flexible UPD approach for synthesis of mono- and bi-metallic Cu, Fe, Co, and Ni SACs directly on oxidized, commercially available carbon electrodes is reported. The UPD mechanism is investigated using in situ X-ray absorption spectroscopy and, finally, the catalytic performance of a UPD-synthesized Co SAC is assessed for electrochemical nitrate reduction to ammonia. The findings expand upon the usefulness and versatility of UPD for SAC synthesis, with hopes of enabling future research toward realization of fast, reliable, and fully electrified SAC synthesis processes., (© 2024 Wiley‐VCH GmbH.)
- Published
- 2024
- Full Text
- View/download PDF
7. Fabrication of Insoluble Elastin by Enzyme-Free Cross-Linking.
- Author
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Hedtke T, Mende M, Steenbock H, Brinckmann J, Menzel M, Hoehenwarter W, Pietzsch M, Groth T, and Schmelzer CEH
- Subjects
- Amino Acids, Proteolysis, Elastin chemistry, Tropoelastin chemistry
- Abstract
Elastin is an essential extracellular matrix protein that enables tissues and organs such as arteries, lungs, and skin, which undergo continuous deformation, to stretch and recoil. Here, an approach to fabricating artificial elastin with close-to-native molecular and mechanical characteristics is described. Recombinantly produced tropoelastin are polymerized through coacervation and allysine-mediated cross-linking induced by pyrroloquinoline quinone (PQQ). A technique that allows the recovery and repeated use of PQQ for protein cross-linking by covalent attachment to magnetic Sepharose beads is developed. The produced material closely resembles natural elastin in its molecular, biochemical, and mechanical properties, enabled by the occurrence of the cross-linking amino acids desmosine, isodesmosine, and merodesmosine. It possesses elevated resistance against tryptic proteolysis, and its Young's modulus ranging between 1 and 2 MPa is similar to that of natural elastin. The approach described herein enables the engineering of mechanically resilient, elastin-like materials for biomedical applications., (© 2023 The Authors. Macromolecular Bioscience published by Wiley-VCH GmbH.)
- Published
- 2023
- Full Text
- View/download PDF
8. Contrasting Capability of Single Atom Palladium for Thermocatalytic versus Electrocatalytic Nitrate Reduction Reaction.
- Author
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Wu X, Nazemi M, Gupta S, Chismar A, Hong K, Jacobs H, Zhang W, Rigby K, Hedtke T, Wang Q, Stavitski E, Wong MS, Muhich C, and Kim JH
- Abstract
The occurrence of high concentrations of nitrate in various water resources is a significant environmental and human health threat, demanding effective removal technologies. Single atom alloys (SAAs) have emerged as a promising bimetallic material architecture in various thermocatalytic and electrocatalytic schemes including nitrate reduction reaction (NRR). This study suggests that there exists a stark contrast between thermocatalytic (T-NRR) and electrocatalytic (E-NRR) pathways that resulted in dramatic differences in SAA performances. Among Pd/Cu nanoalloys with varying Pd-Cu ratios from 1:100 to 100:1, Pd/Cu
(1:100) SAA exhibited the greatest activity (TOFPd = 2 min-1 ) and highest N2 selectivity (94%) for E-NRR, while the same SAA performed poorly for T-NRR as compared to other nanoalloy counterparts. DFT calculations demonstrate that the improved performance and N2 selectivity of Pd/Cu(1:100) in E-NRR compared to T-NRR originate from the higher stability of NO3 * in electrocatalysis and a lower N2 formation barrier than NH due to localized pH effects and the ability to extract protons from water. This study establishes the performance and mechanistic differences of SAA and nanoalloys for T-NRR versus E-NRR., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)- Published
- 2023
- Full Text
- View/download PDF
9. Catalytic Membrane with Copper Single-Atom Catalysts for Effective Hydrogen Peroxide Activation and Pollutant Destruction.
- Author
-
Ma W, Sun M, Huang D, Chu C, Hedtke T, Wang X, Zhao Y, Kim JH, and Elimelech M
- Subjects
- Copper, Peroxides, Sulfhydryl Compounds, Sulfur, Water, Environmental Pollutants, Hydrogen Peroxide
- Abstract
The superior catalytic property of single-atom catalysts (SACs) renders them highly desirable in the energy and environmental fields. However, using SACs for water decontamination is hindered by their limited spatial distribution and density on engineered surfaces and low stability in complex aqueous environments. Herein, we present copper SACs (Cu
1 ) anchored on a thiol-doped reactive membrane for water purification. We demonstrate that the fabricated Cu1 features a Cu-S2 coordination─one copper atom is bridged by two thiolate sulfur atoms, resulting in high-density Cu-SACs on the membrane (2.1 ± 0.3 Cu atoms per nm2 ). The Cu-SACs activate peroxide to generate hydroxyl radicals, exhibiting fast kinetics, which are 40-fold higher than those of nanoparticulate Cu catalysts. The Cu1 -functionalized membrane oxidatively removes organic pollutants from feedwater in the presence of peroxide, achieving efficient water purification. We provide evidence that a dual-site cascade mechanism is responsible for in situ regeneration of Cu1 . Specifically, one of the two linked sulfur atoms detaches the oxidized Cu1 while donating one electron, and an adjacent free thiol rebinds the reduced Cu(I)-S pair, retrieving the Cu-S2 coordination on the reactive membrane. This work presents a universal, facile approach for engineering robust SACs on water-treatment membranes and broadens the application of SACs to real-world environmental problems.- Published
- 2022
- Full Text
- View/download PDF
10. Single-Atom Cobalt Incorporated in a 2D Graphene Oxide Membrane for Catalytic Pollutant Degradation.
- Author
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Wu X, Rigby K, Huang D, Hedtke T, Wang X, Chung MW, Weon S, Stavitski E, and Kim JH
- Subjects
- Catalysis, Cobalt chemistry, Environmental Pollutants, Graphite
- Abstract
We introduce a new graphene oxide (GO)-based membrane architecture that hosts cobalt catalysts within its nanoscale pore walls. Such an architecture would not be possible with catalysts in nanoscale, the current benchmark, since they would block the pores or alter the pore structure. Therefore, we developed a new synthesis procedure to load cobalt in an atomically dispersed fashion, the theoretical limit in material downsizing. The use of vitamin C as a mild reducing agent was critical to load Co as dispersed atoms (Co
1 ), preserving the well-stacked 2D structure of GO layers. With the addition of peroxymonosulfate (PMS), the Co1 -GO membrane efficiently degraded 1,4-dioxane, a small, neutral pollutant that passes through nanopores in single-pass treatment. The observed 1,4-dioxane degradation kinetics were much faster (>640 times) than the kinetics in suspension and the highest among reported persulfate-based 1,4-dioxane destruction. The capability of the membrane to reject large organic molecules alleviated their effects on radical scavenging. Furthermore, the advanced oxidation also mitigated membrane fouling. The findings of this study present a critical advance toward developing catalytic membranes with which two distinctive and complementary processes, membrane filtration and advanced oxidation, can be combined into a single-step treatment.- Published
- 2022
- Full Text
- View/download PDF
11. Engineered Nanoconfinement Accelerating Spontaneous Manganese-Catalyzed Degradation of Organic Contaminants.
- Author
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Zhang S, Hedtke T, Wang L, Wang X, Cao T, Elimelech M, and Kim JH
- Subjects
- Catalysis, Manganese, Oxidation-Reduction, Oxides, Water Pollutants, Chemical, Water Purification
- Abstract
Manganese(III/IV) oxide minerals are known to spontaneously degrade organic pollutants in nature. However, the kinetics are too slow to be useful for engineered water treatment processes. Herein, we demonstrate that nanoscale Mn
3 O4 particles under nanoscale spatial confinement (down to 3-5 nm) can significantly accelerate the kinetics of pollutant degradation, nearly 3 orders of magnitude faster compared to the same reaction in the unconfined bulk phase. We first employed an anodized aluminum oxide scaffold with uniform channel dimensions for experimental and computational studies. We found that the observed kinetic enhancement resulted from the increased surface area of catalysts exposed to the reaction, as well as the increased local proton concentration at the Mn3 O4 surface and subsequent acceleration of acid-catalyzed reactions even at neutral pH in bulk. We further demonstrate that a reactive Mn3 O4 -functionalized ceramic ultrafiltration membrane, a more suitable scaffold for realistic water treatment, achieved nearly complete removal of various phenolic and aniline pollutants, operated under a common ultrafiltration water flux. Our findings mark an important advance toward the development of catalytic membranes that can degrade pollutants in addition to their intrinsic function as a physical separation barrier, especially since they are based on accelerating natural catalytic pathways that do not require any chemical addition.- Published
- 2021
- Full Text
- View/download PDF
12. Membrane-Confined Iron Oxychloride Nanocatalysts for Highly Efficient Heterogeneous Fenton Water Treatment.
- Author
-
Zhang S, Hedtke T, Zhu Q, Sun M, Weon S, Zhao Y, Stavitski E, Elimelech M, and Kim JH
- Subjects
- Hydrogen Peroxide, Hydroxyl Radical, Oxidation-Reduction, Iron Compounds, Water Pollutants, Chemical, Water Purification
- Abstract
Heterogeneous advanced oxidation processes (AOPs) allow for the destruction of aqueous organic pollutants via oxidation by hydroxyl radicals (
• OH). However, practical treatment scenarios suffer from the low availability of short-lived• OH in aqueous bulk, due to both mass transfer limitations and quenching by water constituents, such as natural organic matter (NOM). Herein, we overcome these challenges by loading iron oxychloride catalysts within the pores of a ceramic ultrafiltration membrane, resulting in an internal heterogeneous Fenton reaction that can degrade organics in complex water matrices with pH up to 6.2. With• OH confined inside the nanopores (∼ 20 nm), this membrane reactor completely removed various organic pollutants with water fluxes of up to 100 L m-2 h-1 (equivalent to a retention time of 10 s). This membrane, with a pore size that excludes NOM (>300 kDa), selectively exposed smaller organics to• OH within the pores under confinement and showed excellent resiliency to representative water matrices (simulated surface water and sand filtration effluent samples). Moreover, the membrane exhibited sustained AOPs (>24 h) and could be regenerated for multiple cycles. Our results suggest the feasibility of exploiting ultrafiltration membrane-based AOP platforms for organic pollutant degradation in complex water scenarios.- Published
- 2021
- Full Text
- View/download PDF
13. Towards the Therapeutic Use of Thrombospondin 1/CD47 Targeting TAX2 Peptide as an Antithrombotic Agent.
- Author
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Jeanne A, Sarazin T, Charlé M, Kawecki C, Kauskot A, Hedtke T, Schmelzer CEH, Martiny L, Maurice P, and Dedieu S
- Subjects
- Animals, Arterial Occlusive Diseases blood, Arterial Occlusive Diseases metabolism, CD47 Antigen metabolism, Collagen metabolism, Disease Models, Animal, Fibrinolytic Agents toxicity, Humans, Male, Mice, Inbred C57BL, Mice, Knockout, Peptides, Cyclic toxicity, Platelet Aggregation Inhibitors toxicity, Rats, Sprague-Dawley, Signal Transduction, Thrombosis blood, Thrombosis metabolism, Thrombospondin 1 genetics, Thrombospondin 1 metabolism, Time Factors, Mice, Rats, Arterial Occlusive Diseases prevention & control, CD47 Antigen antagonists & inhibitors, Fibrinolytic Agents pharmacology, Peptides, Cyclic pharmacology, Platelet Aggregation drug effects, Platelet Aggregation Inhibitors pharmacology, Thrombosis prevention & control, Thrombospondin 1 antagonists & inhibitors
- Abstract
Objective: TSP-1 (thrombospondin 1) is one of the most expressed proteins in platelet α-granules and plays an important role in the regulation of hemostasis and thrombosis. Interaction of released TSP-1 with CD47 membrane receptor has been shown to regulate major events leading to thrombus formation, such as, platelet adhesion to vascular endothelium, nitric oxide/cGMP (cyclic guanosine monophosphate) signaling, platelet activation as well as aggregation. Therefore, targeting TSP-1:CD47 axis may represent a promising antithrombotic strategy. Approach and Results: A CD47-derived cyclic peptide was engineered, namely TAX2, that targets TSP-1 and selectively prevents TSP-1:CD47 interaction. Here, we demonstrate for the first time that TAX2 peptide strongly decreases platelet aggregation and interaction with collagen under arterial shear conditions. TAX2 also delays time for complete thrombotic occlusion in 2 mouse models of arterial thrombosis following chemical injury, while Thbs1
-/- mice recapitulate TAX2 effects. Importantly, TAX2 administration is not associated with increased bleeding risk or modification of hematologic parameters., Conclusions: Overall, this study sheds light on the major contribution of TSP-1:CD47 interaction in platelet activation and thrombus formation while putting forward TAX2 as an innovative antithrombotic agent with high added-value.- Published
- 2021
- Full Text
- View/download PDF
14. Direct three-dimensional imaging for morphological analysis of electrospun fibers with laboratory-based Zernike X-ray phase-contrast computed tomography.
- Author
-
Santos de Oliveira C, González AT, Hedtke T, Kürbitz T, Heilmann A, Schmelzer CEH, and Martins de S E Silva J
- Subjects
- Imaging, Three-Dimensional, Nanofibers chemistry, Particle Size, Porosity, Gelatin chemistry, Tomography, X-Ray Computed methods
- Abstract
Electrospinning is a well-established and widely used method for the production of protein-based fibrous biomaterials. The visualization of the morphology and the characterization of sample features related to the three-dimensional (3D) structure, like the porosity and fibers thickness, is crucial for the design and fabrication of tailor-made and application-optimized materials. Here, we evaluated the benefits of using 3D X-ray imaging in a laboratory setup with a resolution in the sub-micrometer range for the characterization of electrospun gelatin fibrous mats. We used phase-contrast X-ray computed tomography at the nanoscale (nano-CT) for the evaluation of the time-course morphological changes of the mats induced by chemical cross-linking of the gelatin fibers. We present an image processing protocol that enables the segmentation of the fibers and quantification of the mats porosity, the analysis of the shape and size of the pores, and of the fibers thickness and orientation. We compared the results obtained from the processed nano-CT data with those obtained with the conventional methods used for the characterization of electrospun fibrous materials, and we discuss the advantages and limitations of each method when applied to gelatin electrospun samples. Our results reveal that the use of phase-contrast nano-CT provides quick additional and relevant information for the characterization of fibrous mats and, thus, provides beneficial insights for the design and fabrication of novel fibrous materials., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
15. Mechanism of Heterogeneous Fenton Reaction Kinetics Enhancement under Nanoscale Spatial Confinement.
- Author
-
Zhang S, Sun M, Hedtke T, Deshmukh A, Zhou X, Weon S, Elimelech M, and Kim JH
- Subjects
- Catalysis, Kinetics, Oxidation-Reduction, Hydrogen Peroxide, Hydroxyl Radical
- Abstract
Nanoscale catalysts that can enable Fenton-like chemistry and produce reactive radicals from hydrogen peroxide activation have been extensively studied in order to overcome the limitations of homogeneous Fenton processes. Despite several advantageous features, limitation in mass transfer of short-lived radical species is an inherent drawback of the heterogeneous system. Here, we present a mechanistic foundation for the way spatial confinement of Fenton chemistry at the nanoscale can significantly enhance the kinetics of radical-mediated oxidation reactions-pollutant degradation in particular. We synthesized a series of Fe
3 O4 -functionalized nanoreactors with precise pore dimensions, based on an anodized aluminum oxide template, to enable quantitative analysis of nanoconfinement effects. Combined with computational simulation of spatial distribution of radicals, we found that hydroxyl radical concentration was strongly dependent on the distance from the surface of Fenton catalysts. This distance dependency significantly influences the gross reaction kinetics and accounts for the observed nanoconfinement effects. We further found that a length scale below 25 nm is critical to avoid the limitation of short-lived species diffusion and achieve kinetics that are orders of magnitude faster than those obtained in a batch suspension of heterogeneous catalysts. These findings suggest a new strategy to develop an innovative heterogeneous catalytic system with the most effective use of hydroxyl radicals in oxidation treatment scenarios.- Published
- 2020
- Full Text
- View/download PDF
16. Unique molecular networks: Formation and role of elastin cross-links.
- Author
-
Schmelzer CEH, Hedtke T, and Heinz A
- Subjects
- 2-Aminoadipic Acid analogs & derivatives, 2-Aminoadipic Acid metabolism, Animals, Blood Vessels chemistry, Blood Vessels metabolism, Connective Tissue Diseases pathology, Elastic Tissue chemistry, Elastin chemistry, Humans, Ligaments chemistry, Ligaments metabolism, Lung chemistry, Lung metabolism, Lysine metabolism, Microfibrils chemistry, Microfibrils metabolism, Oxidation-Reduction, Skin chemistry, Skin metabolism, Aging metabolism, Connective Tissue Diseases metabolism, Elastic Tissue metabolism, Elastin metabolism, Protein Processing, Post-Translational, Protein-Lysine 6-Oxidase metabolism
- Abstract
Elastic fibers are essential assemblies of vertebrates and confer elasticity and resilience to various organs including blood vessels, lungs, skin, and ligaments. Mature fibers, which comprise a dense and insoluble elastin core and a microfibrillar mantle, are extremely resistant toward intrinsic and extrinsic influences and maintain elastic function over the human lifespan in healthy conditions. The oxidative deamination of peptidyl lysine to peptidyl allysine in elastin's precursor tropoelastin is a crucial posttranslational step in their formation. The modification is catalyzed by members of the family of lysyl oxidases and the starting point for subsequent manifold condensation reactions that eventually lead to the highly cross-linked elastomer. This review summarizes the current understanding of the formation of cross-links within and between the monomer molecules, the molecular sites, and cross-link types involved and the pathological consequences of abnormalities in the cross-linking process., (© 2019 The Authors. IUBMB Life published by Wiley Periodicals, Inc. on behalf of International Union of Biochemistry and Molecular Biology.)
- Published
- 2020
- Full Text
- View/download PDF
17. Expression of elastolytic cathepsins in human skin and their involvement in age-dependent elastin degradation.
- Author
-
Panwar P, Hedtke T, Heinz A, Andrault PM, Hoehenwarter W, Granville DJ, Schmelzer CEH, and Brömme D
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Cathepsin K analysis, Cathepsins analysis, Child, Cysteine Endopeptidases analysis, Elastin analysis, Elastin ultrastructure, Female, Humans, Middle Aged, Proteolysis, Young Adult, Cathepsin K metabolism, Cathepsins metabolism, Cysteine Endopeptidases metabolism, Elastin metabolism, Skin metabolism, Skin Aging
- Abstract
Background: Skin ageing is associated with structure-functional changes in the extracellular matrix, which is in part caused by proteolytic degradation. Since cysteine cathepsins are major matrix protein-degrading proteases, we investigated the age-dependent expression of elastolytic cathepsins K, S, and V in human skin, their in vitro impact on the integrity of the elastic fibre network, their cleavage specificities, and the release of bioactive peptides., Methods: Cathepsin-mediated degradation of human skin elastin samples was assessed from young to very old human donors using immunohistochemical and biochemical assays, scanning electron microscopy, and mass spectrometry., Results: Elastin samples derived from patients between 10 and 86 years of age were analysed and showed an age-dependent deterioration of the fibre structure from a dense network of thinner fibrils into a beaded and porous mesh. Reduced levels of cathepsins K, S, and V were observed in aged skin with a predominant epidermal expression. Cathepsin V was the most potent elastase followed by cathepsin K and S. Biomechanical analysis of degraded elastin fibres corroborated the destructive activity of cathepsins. Mass spectrometric determination of the cleavage sites in elastin revealed that all three cathepsins predominantly cleaved in hydrophobic domains. The degradation of elastin was efficiently inhibited by an ectosteric inhibitor. Furthermore, the degradation of elastin fibres resulted in the release of bioactive peptides, which have previously been associated with various pathologies., Conclusion: Cathepsins are powerful elastin-degrading enzymes and capable of generating a multitude of elastokines. They may represent a viable target for intervention strategies to reduce skin ageing., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
18. A comprehensive map of human elastin cross-linking during elastogenesis.
- Author
-
Hedtke T, Schräder CU, Heinz A, Hoehenwarter W, Brinckmann J, Groth T, and Schmelzer CEH
- Subjects
- Amino Acid Sequence genetics, Desmosine chemistry, Elastic Tissue chemistry, Elastic Tissue ultrastructure, Elastin ultrastructure, Extracellular Matrix chemistry, Extracellular Matrix ultrastructure, Humans, Hydrophobic and Hydrophilic Interactions, Mass Spectrometry, Molecular Structure, Protein Isoforms chemistry, Protein Isoforms ultrastructure, Protein Processing, Post-Translational genetics, Skin chemistry, Tropoelastin ultrastructure, Elastin chemistry, Lysine chemistry, Peptides chemistry, Tropoelastin chemistry
- Abstract
Elastin is an essential structural protein in the extracellular matrix of vertebrates. It is the core component of elastic fibers, which enable connective tissues such as those of the skin, lungs or blood vessels to stretch and recoil. This function is provided by elastin's exceptional properties, which mainly derive from a unique covalent cross-linking between hydrophilic lysine-rich motifs of units of the monomeric precursor tropoelastin. To date, elastin's cross-linking is poorly investigated. Here, we purified elastin from human tissue and cleaved it into soluble peptides using proteases with different specificities. We then analyzed elastin's molecular structure by identifying unmodified residues, post-translational modifications and cross-linked peptides by high-resolution mass spectrometry and amino acid analysis. The data revealed the presence of multiple isoforms in parallel and a complex and heterogeneous molecular interconnection. We discovered that the same lysine residues in different monomers were simultaneously involved in various cross-link types or remained unmodified. Furthermore, both types of cross-linking domains, Lys-Pro and Lys-Ala domains, participate not only in bifunctional inter- but also in intra-domain cross-links. We elucidated the sequences of several desmosine-containing peptides and the contribution of distinct domains such as 6, 14 and 25. In contrast to earlier assumptions proposing that desmosine cross-links are formed solely between two domains, we elucidated the structure of a peptide that proves a desmosine formation with participation of three Lys-Ala domains. In summary, these results provide new and detailed insights into the cross-linking process, which takes place within and between human tropoelastin units in a stochastic manner., (© 2019 Federation of European Biochemical Societies.)
- Published
- 2019
- Full Text
- View/download PDF
19. Lysyl oxidase-like 2 (LOXL2)-mediated cross-linking of tropoelastin.
- Author
-
Schmelzer CEH, Heinz A, Troilo H, Lockhart-Cairns MP, Jowitt TA, Marchand MF, Bidault L, Bignon M, Hedtke T, Barret A, McConnell JC, Sherratt MJ, Germain S, Hulmes DJS, Baldock C, and Muller L
- Subjects
- Animals, CHO Cells, Catalytic Domain physiology, Cell Line, Collagen metabolism, Cricetulus, Elastin metabolism, Extracellular Matrix metabolism, Humans, Proteolysis, Substrate Specificity physiology, Amino Acid Oxidoreductases metabolism, Tropoelastin metabolism
- Abstract
Lysyl oxidases (LOXs) play a central role in extracellular matrix remodeling during development and tumor growth and fibrosis through cross-linking of collagens and elastin. We have limited knowledge of the structure and substrate specificity of these secreted enzymes. LOXs share a conserved C-terminal catalytic domain but differ in their N-terminal region, which is composed of 4 repeats of scavenger receptor cysteine-rich (SRCR) domains in LOX-like (LOXL) 2. We investigated by X-ray scattering and electron microscopy the low-resolution structure of the full-length enzyme and the structure of a shorter form lacking the catalytic domain. Our data demonstrate that LOXL2 has a rod-like structure with a stalk composed of the SRCR domains and the catalytic domain at its tip. We detected direct interaction between LOXL2 and tropoelastin (TE) and also LOXL2-mediated deamination of TE. Using proteomics, we identified several allysines together with cross-linked TE peptides. The elastin-like material generated was resistant to trypsin proteolysis and displayed mechanical properties similar to mature elastin. Finally, we detected the codistribution of LOXL2 and elastin in the vascular wall. Altogether, these data suggest that LOXL2 could participate in elastogenesis in vivo and could be used as a means of cross-linking TE in vitro for biomimetic and cell-compatible tissue engineering purposes.-Schmelzer, C. E. H., Heinz, A., Troilo, H., Lockhart-Cairns, M.-P., Jowitt, T. A., Marchand, M. F., Bidault, L., Bignon, M., Hedtke, T., Barret, A., McConnell, J. C., Sherratt, M. J., Germain, S., Hulmes, D. J. S., Baldock, C., Muller, L. Lysyl oxidase-like 2 (LOXL2)-mediated cross-linking of tropoelastin.
- Published
- 2019
- Full Text
- View/download PDF
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