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Canis Major OB1 (CMa OB1) is a Galactic stellar association with a very intriguing star-formation scenario. There are more than two dozen known star clusters in its line of sight, but it is not clear which ones are physically associated with CMa OB1. We use a clustering code that employs 5-dimensional data from the Gaia DR2 catalogue to identify physical groups and obtain their astrometric parameters and, in addition, we use two different isochrone-fitting methods to estimate the ages of these groups. We find 15 stellar groups with distances between 570 pc and 1650 pc, including 10 previously known and 5 new open cluster candidates. Four groups, precisely the youngest ones ($, Comment: 23 pages, 28 figures, 7 tables

The relationship between young stellar clusters and respective parental molecular clouds is still an open issue: for instance, are the similarities between substructures of clouds and clusters just a coincidence? Or would they be the indication of a physical relationship? In order to address these issues, we have studied the CMa OB1/R1 region that shows evidence for a complex star formation history. We obtained molecular clouds mapping with the IRAM-30 metre telescope to reveal the physical conditions of an unexplored side of the CMa region aiming to compare the morphology of the clouds with the distribution of the young stellar objects (YSOs). We also study the clouds kinematics searching for gradients and jet signatures that could trace different star formation scenarios. The YSOs were selected on the basis of astrometric data from Gaia EDR3 that characterise the moving groups. The distance of 1099$_{-24}^{+25}$ pc was obtained for the sample, based on the mean error-weighted parallax. Optical and near-infrared photometry is used to verify the evolutionary status and circumstellar characteristics of the YSOs. Among the selected candidates we found 40 members associated with the cloud: 1 Class I, 11 Class II, and 28 Class III objects. Comparing the spatial distribution of the stellar population with the cores revealed by the 13CO map, we verify that peaks of emission coincide with the position of YSOs confirming the association of these objects to their dense natal gas. Our observations support the large-scale scenario of the CMa shell-like structure formed as a relic of successive supernova events., A&A (accepted): 11 pages with 8 figures and 4 Tables

The scenario of star cluster formation can be better understood based on the detailed study of the dynamical conditions of the associated gas, clustering properties and effects of ionizing sources, among others. Some of these characteristics are explored in our ongoing work based on observations with the SOAR and T80-S telescopes. With these data we have obtained a complete multi-band photometric catalogue of selected clusters that we characterize using color-color diagrams and flux ratios. In particular, this work is focused on SOAR/Spartan (near-infrared) observations of the Canis Major star-forming region.

Investigating the proteolytic activity of the recombinant Mycobacterium leprae Heat Shock Protein of 65 kDa (rHsp65), chaperonin 2 (cpn2), we observed that it displays high instability. The fragmentation process starts at the C-terminus followed by progressive degradation of the N-terminus, which leads to a stable fragment comprising the middle region of the molecule. Urea was able to prevent autolysis, probably due to its denaturing action, while EDTA increased degradation levels indicating the need for metal ions. Peptides originated from autolysis were purified and analyzed by mass spectrometry, generating a continuous map. Since the bacteria and mammalian Hsp60 are known to be targets of the immune response and have been implicated in autoimmune diseases and chronic inflammation, the in vivo effect of rHsp65 peptides was evaluated in the spontaneous Systemic Lupus Erythematosus (SLE) model developed by the (NZB/NZW)F1 mouse hybrids, and their individual anti-rHsp65 IgG2a/IgG1 antibody titer ratio was determined. The results showed orientation toward a TH1 responsiveness, and the treatment with the rHsp65 peptides diminished the environmental variance of the survival time of treated animals. These results outline the fact that environmental factors may also act through the modified stability expression of Heat Shock Proteins intervening during autoimmune processes.

The bradykinin-potentiating peptides from Bothrops jararaca venom are the most potent natural inhibitors of the angiotensin-converting enzyme. The biochemical and biological features of these peptides were crucial to demonstrate the pivotal role of the angiotensin-converting enzyme in blood pressure regulation. In the present study, seven bradykinin-potentiating peptides were identified within the C-type natriuretic peptide precursor cloned from snake brain. The bradykinin-potentiating peptides deduced from the B. jararaca brain precursor are strong in vitro inhibitors of the angiotensin-converting enzyme (nanomolar range), and also potentiate the bradykinin effects in ex vivo and in vivo experiments. Two of these peptides are novel bradykinin-potentiating peptides, one of which displays high specificity toward the N-domain active site of the somatic angiotensin-converting enzyme. In situ hybridization studies revealed the presence of the bradykinin-potentiating peptides precursor mRNAs in distinct regions of the B. jararaca brain, such as the ventromedial hypothalamus, the paraventricular nuclei, the paraventricular organ, and the subcommissural organ. The biochemical and pharmacological properties of the brain bradykinin-potentiating peptides, their presence within the neuroendocrine regulator C-type natriuretic peptide precursor, and their expression in regions of the snake brain correlated to neuroendocrine functions, strongly suggest that these peptides belong to a novel class of endogenous vasoactive peptides.

Aiming to contribute to the understanding of star formation and evolution in the Canis Major (CMa R1) Molecular Clouds Complex, we analyze the spectral characteristics of a population of young stars associated with the arc-shaped nebula Sh2-296. Our XMM/Newton observations detected 109 X-ray sources in the region and optical spectroscopy was performed with Gemini telescope for 85 optical counterparts. We identified and characterized 51 objects that present features typically found in young objects, such as Hα emission and strong absorption on the Li I line.

Lonomia obliqua venom causes a severe consumptive coagulopathy, which can lead to a hemorrhagic syndrome. The crude bristles extract displays a procoagulant activity due to a Factor X and to a prothrombin activating activity. Here, we describe a 69 kDa prothrombin activator serine protease purified from L. obliqua caterpillar bristle extract using gel filtration (Sephadex G 75) and HPLC (C 4 column). The purified protein was able to activate prothrombin in a dose-dependent manner, and calcium ions increased this activity. The prothrombin-derived fluorogenic peptide (Abz-YQTFFNPRTGSQ-EDDnp) had its main cleavage site at the Arg–Thr bond. The kinetic parameters obtained for this substrate were K m app of 4.5 μM, k cat of 5.32 s −1 , and a k cat / K m app of 1.2×10 6 M −1 s −1 . The prothrombin fragments generated by the purified enzyme corresponded to the molecular masses of prethrombin 2, fragment 1, fragment 2, and thrombin as seen in SDS-PAGE. The thrombin generated was able to clot purified fibrinogen. The partial amino acid sequence of the purified protein, named Lopap ( L. obliqua prothrombin activator protease), showed no similarity to any known prothrombin activator.

Increasing occurrence of hemorrhagic syndrome in man, caused by contact with Lonomia obliqua caterpillars, has been reported in Southern Brazil in the past 10 years. The L. obliqua venom causes a severe consumptive coagulopathy, which can lead to hemorrhagic syndrome. L. obliqua prothrombin activator protease (Lopap) is a 69-kDa prothrombin activator serine protease isolated from L. obliqua caterpillar bristle extract, which is able to evoke thrombus formation, unclottable blood, and fibrinogen depletion when injected into the blood stream of rats. The purified protein generated thrombin from prothrombin, able to clot purified human fibrinogen and plasma. A decrease in platelet count and inhibition of collagen-induced platelet aggregation were observed, as well as leukocyte infiltration in the lungs. In addition, we observed congestion and hemorrhage in renal glomeruli and necrosis in renal distal tubules. These data support the hypothesis that Lopap contributes to the clinical syndrome caused by human contact with L. obliqua, most probably through prothrombin activation, resulting in a consumption coagulopathy.

A indústria automotiva responde, atualmente, por uma grande parte do mercado de consumo de plásticos, por isso existe um crescente interesse no investimento em processos de reciclagem, inclusive devido à vinculação às atividades relacionadas à proteção ambiental. Dentre as peças automotivas, os pára-choques são relativamente fáceis de serem reciclados devido às suas dimensões e por constituírem, geralmente, de um único material, o polipropileno (PP). Neste trabalho, analisaram-se as propriedades mecânicas de misturas de PP virgem e reciclado, em três porcentagens diferentes, a fim de detectar as alterações das mesmas, relacionando-as à qualidade do produto. Concluiu-se que porcentagens de reciclado acima de 30% causam deterioração das propriedades mecânicas do produto. Este estudo visa a fornecer uma contribuição para o aumento na utilização de plásticos reciclados na indústria automotiva.The automotive industry is now responsible for a large share of plastic consumption, which prompted increasing investments in recycling processes, also due to the need of ambient protection. Among the automotives parts, bumpers are relatively easy to be recycled due to their dimensions and because they are made of a single material, polypropylene (PP). In this work, the mechanical properties of mixtures of virgin and recycled PP, in three different percentages, were analyzed in order to detect alterations and associate them with the product quality. It was concluded that percentages of recycled material above 30% deteriorate the mechanical properties of the product. This study aims to provide a contribution to the increase in the recycled plastic use in the automotive industry.

The widely used plasmid pUC19 transformed Proteus mirabilis with the high efficiency of 3−7 × 105 transformants/μg plasmid DNA, when a modification of the electroporation method was employed. Moreover, the Bacillus subtilis α-amylase gene was expressed in P. mirabilis upon introduction of the pUC19 recombinant vector, as could be demonstrated by the occurrence of starch-degradation halos surrounding transformant colonies and starch consumption in liquid culture supernatants.

The recombinant rat testes metallo-endooligopeptidase (EC 3.4.24.15) and the rabbit brain endooligopeptidase A (formerly EC 3.4.22.19) were compared, side-by-side, in view of their striking similarities in both the physicochemical features and the specificities for oligopeptides. Concerning the tissue distribution in rat and rabbit, no relation between the levels of enzyme activity in cytosol and the levels of metallo-endooligopeptidase 24.15 mRNA could be established. The results suggest that the predominant neuropeptide-metabolizing activity attributed to the metallo-endooligopeptidase 24.15 is performed by, at least, two distinct cytosolic enzymes, one predominant in rat testes and the other in rabbit brain and testes, and possibly also in rat brain. Both enzymes are activated by dithiothreitol and irreversibly inhibited by a SH-affinity labeling dynorphin-related compound, but they are not inhibited by EDTA in a concentration dependent manner. Both enzymes exhibit the same specificity toward several bioactive peptides, except for LH-RH and substance P, which are only hydrolysed by the rat testes enzyme. Taken together, these results lead us to conclude that it is unlikely that the recombinant rat testes metallo-endooligopeptidase 24.15 and the rabbit brain endooligopeptidase A are the same molecule although they might belong to the same family of oligopeptidases.

A series of biologically active peptides and related compounds (opioid peptides, neurotensin, and bradykinin) were used as substrates or competitive inhibitors to study the structural requirements for peptide interaction with endopeptidase 22.19. The kinetics of hydrolysis of these peptides indicated that, in contrast to other proteases, the substrate specificity of endopeptidase 22.19 is not determined by the amino acids flanking the sensitive bonds of the substrates. The competition between bioactive peptide analogues and the quenched fluorescence substrate of endopeptidase 22.19 indicated that their length and their flexibility may be the dominant factors to explain their binding specificities. These peculiar features of endopeptidase 22.19 may be of importance to understand the physiological processes of conversion and inactivation of biologically active peptides.

Hsp60 is an abundant and highly conserved family of intracellular molecules. Increased levels of this family of proteins have been observed in the extracellular compartment in chronic inflammation. Administration of M. leprae Hsp65 [WT] in [NZBxNZW]F(1) mice accelerates the Systemic Lupus Erythematosus [SLE] progression whereas the point mutated K(409)A Hsp65 protein delays the disease. Here, the biological effects of M. leprae Hsp65 Leader pep and K(409)A pep synthetic peptides, which cover residues 352-371, are presented. Peptides had immunomodulatory effects similar to that observed with their respective proteins on survival and the combined administration of K(409)A+Leader pep or K(409)A pep+WT showed that the mutant forms were able to inhibit the deleterious effect of WT on mortality, indicating the neutralizing potential of the mutant molecules in SLE progression. Molecular modeling showed that replacing Lysine by Alanine affects the electrostatic potential of the 352-371 region. The number of interactions observed for WT is much higher than for Hsp65 K(409)A and mouse Hsp60. The immunomodulatory effects of the point-mutated protein and peptide occurred regardless of the catalytic activity. These findings may be related to the lack of effect on survival when F(1) mice were inoculated with Hsp60 or K(409)A pep. Our findings indicate the use of point-mutated Hsp65 molecules, such as the K(409)A protein and its corresponding peptide, that may minimize or delay the onset of SLE, representing a new approach to the treatment of autoimmune diseases.

BPPs have been identified in the venom of the Bothrops jararaca snake, or deduced from precursor proteins expressed either in the venom gland or in the brain of the snake. Their potentiating activity on bradykinin (Bk) is assumed to occur through a somatic angiotensin-converting enzyme (sACE) inhibitory mechanism. We have demonstrated that synthetic BPPs show remarkable functional differences, despite their high amino acid sequence similarities. Recently, we demonstrated that BPP-10c, after i.p. administration, was found in its intact form and in the form of a unique metabolite (des-Pro10 BPP-10c) in mouse urine. Given this finding, we selected a number of BPPs with different structure-activities – BPP-5a (

The snake venom proline-rich peptide BPP 10c is an active somatic angiotensin-converting enzyme (sACE) inhibitors. Recently we demonstrated that the anti-hypertensive effect of BPP 10c is not related to the inhibition of sACE alone, thus suggesting that this enzyme is not its only target for blood pressure reduction. In the present work, a biodistribution study in Swiss mice of [125I]-BPP 10c in the absence or in the presence of a saturating concentration of captopril, a selective active-site inhibitor of sACE, demonstrated that: (1) [125I]-BPP 10c was present in several organs and the renal absorption was significantly high; (2) [125I]-BPP 10c showed a clear preference for the kidney, maintaining a high concentration in this organ in the presence of captopril for at least 3 h; (3) The residual amount of [125I]-BPP 10c in the kidney of animals simultaneously treated with captopril suggest that the peptide can interact with other targets different from sACE in this organ. We also showed that Cy3-labeled BPP 10c was internalized by human embryonic kidney cells (HEK-293T). Taken together, these results suggest that sACE inhibition by captopril affects the tissue distribution of [125I]-BPP 10c and that the anti-hypertensive effects of BPP 10c are not only dependent on sACE inhibition.

The bradykinin-potentiating peptides from Bothrops jararaca venom are the most potent natural inhibitors of the angiotensin-converting enzyme. The biochemical and biological features of these peptides were crucial to demonstrate the pivotal role of the angiotensin-converting enzyme in blood pressure regulation. In the present study, seven bradykinin-potentiating peptides were identified within the C-type natriuretic peptide precursor cloned from snake brain. The bradykinin-potentiating peptides deduced from the B. jararaca brain precursor are strong in vitro inhibitors of the angiotensin-converting enzyme (nanomolar range), and also potentiate the bradykinin effects in ex vivo and in vivo experiments. Two of these peptides are novel bradykinin-potentiating peptides, one of which displays high specificity toward the N-domain active site of the somatic angiotensin-converting enzyme. In situ hybridization studies revealed the presence of the bradykinin-potentiating peptides precursor mRNAs in distinct regions of the B. jararaca brain, such as the ventromedial hypothalamus, the paraventricular nuclei, the paraventricular organ, and the subcommissural organ. The biochemical and pharmacological properties of the brain bradykinin-potentiating peptides, their presence within the neuroendocrine regulator C-type natriuretic peptide precursor, and their expression in regions of the snake brain correlated to neuroendocrine functions, strongly suggest that these peptides belong to a novel class of endogenous vasoactive peptides.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:52:39Z No. of bitstreams: 1 S0100-879X2001001100004.pdf: 65223 bytes, checksum: 70eb4858923bff83dbb8b171af5b0ae0 (MD5) Made available in DSpace on 2013-08-22T18:52:39Z (GMT). No. of bitstreams: 1 S0100-879X2001001100004.pdf: 65223 bytes, checksum: 70eb4858923bff83dbb8b171af5b0ae0 (MD5) Previous issue date: 2001-11-01 Made available in DSpace on 2013-09-30T19:44:03Z (GMT). No. of bitstreams: 2 S0100-879X2001001100004.pdf: 65223 bytes, checksum: 70eb4858923bff83dbb8b171af5b0ae0 (MD5) S0100-879X2001001100004.pdf.txt: 25554 bytes, checksum: 06ea4d348f37b61b5cedabdcabfb43c2 (MD5) Previous issue date: 2001-11-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:54:58Z No. of bitstreams: 2 S0100-879X2001001100004.pdf: 65223 bytes, checksum: 70eb4858923bff83dbb8b171af5b0ae0 (MD5) S0100-879X2001001100004.pdf.txt: 25554 bytes, checksum: 06ea4d348f37b61b5cedabdcabfb43c2 (MD5) Made available in DSpace on 2014-05-20T13:54:58Z (GMT). No. of bitstreams: 2 S0100-879X2001001100004.pdf: 65223 bytes, checksum: 70eb4858923bff83dbb8b171af5b0ae0 (MD5) S0100-879X2001001100004.pdf.txt: 25554 bytes, checksum: 06ea4d348f37b61b5cedabdcabfb43c2 (MD5) Previous issue date: 2001-11-01 The opportunistic bacterium Proteus mirabilis secretes a metalloprotease, ZapA, considered to be one of its virulence factors due to its IgA-degrading activity. However, the substrate specificity of this enzyme has not yet been fully characterized. In the present study we used fluorescent peptides derived from bioactive peptides and the oxidized ß-chain of insulin to determine the enzyme specificity. The bradykinin- and dynorphin-derived peptides were cleaved at the single bonds Phe-Ser and Phe-Leu, with catalytic efficiencies of 291 and 13 mM/s, respectively. Besides confirming already published cleavage sites, a novel cleavage site was determined for the ß-chain of insulin (Val-Asn). Both the natural and the recombinant enzyme displayed the same broad specificity, demonstrated by the presence of hydrophobic, hydrophilic, charged and uncharged amino acid residues at the scissile bonds. Native IgA, however, was resistant to hydrolysis by ZapA. Universidade de São Paulo Instituto de Ciências Biomédicas Departamento de Microbiologia Instituto Butantan CEPID-FAPESP Centro de Toxinologia Aplicada Universidade Federal de São Paulo (UNIFESP) CEPID-FAPESP Centro de Toxinologia Aplicada Universidade Estadual Paulista CEPID-FAPESP Centro de Toxinologia Aplicada Universidade Estadual Paulista CEPID-FAPESP Centro de Toxinologia Aplicada

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:52:38Z No. of bitstreams: 1 S0100-879X2000000700006.pdf: 172031 bytes, checksum: cce457ecff6e183722d066458bb8ef73 (MD5) Made available in DSpace on 2013-08-22T18:52:38Z (GMT). No. of bitstreams: 1 S0100-879X2000000700006.pdf: 172031 bytes, checksum: cce457ecff6e183722d066458bb8ef73 (MD5) Previous issue date: 2000-07-01 Made available in DSpace on 2013-09-30T19:44:00Z (GMT). No. of bitstreams: 2 S0100-879X2000000700006.pdf: 172031 bytes, checksum: cce457ecff6e183722d066458bb8ef73 (MD5) S0100-879X2000000700006.pdf.txt: 18445 bytes, checksum: 71add675aa74cd36e8b41f05cff8c168 (MD5) Previous issue date: 2000-07-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:54:58Z No. of bitstreams: 2 S0100-879X2000000700006.pdf: 172031 bytes, checksum: cce457ecff6e183722d066458bb8ef73 (MD5) S0100-879X2000000700006.pdf.txt: 18445 bytes, checksum: 71add675aa74cd36e8b41f05cff8c168 (MD5) Made available in DSpace on 2014-05-20T13:54:58Z (GMT). No. of bitstreams: 2 S0100-879X2000000700006.pdf: 172031 bytes, checksum: cce457ecff6e183722d066458bb8ef73 (MD5) S0100-879X2000000700006.pdf.txt: 18445 bytes, checksum: 71add675aa74cd36e8b41f05cff8c168 (MD5) Previous issue date: 2000-07-01 The protease ZapA, secreted by Proteus mirabilis, has been considered to be a virulence factor of this opportunistic bacterium. The control of its expression requires the use of an appropriate methodology, which until now has not been developed. The present study focused on the replacement of azocasein with fluorogenic substrates, and on the definition of enzyme specificity. Eight fluorogenic substrates were tested, and the peptide Abz-Ala-Phe-Arg-Ser-Ala-Ala-Gln-EDDnp was found to be the most convenient for use as an operational substrate for ZapA. A single peptide bond (Arg-Ser) was cleaved with a Km of 4.6 µM, a k cat of 1.73 s-1, and a catalytic efficiency of 376 (mM s)-1. Another good substrate for ZapA was peptide 6 (Abz-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-Gln-EDDnp) which was cleaved at a single bond (Phe-Ser) with a Km of 13.6 µM, a k cat of 3.96 s-1 and a catalytic efficiency of 291 (mM s)-1. The properties of the amino acids flanking the scissile bonds were also evaluated, and no clear requirement for the amino acid residue at P1 was found, although the enzyme seems to have a preference for a hydrophobic residue at P2. Universidade de São Paulo (USP) Universidade Federal de São Paulo (UNIFESP) Universidade Estadual Paulista (UNESP) Instituto Butantan

In this study we investigated the fate of a class of proteasome-generated oligopeptides, exposing them to the crude cytosol of macrophages or to the purified recombinant thimet oligopeptidase. Among the proteasome products of known sequences are MHC class I epitopes, 13 of which were randomly chosen to be used as putative substrates. Surprisingly, our results clearly showed that the majority of the peptides were poorly or not degraded, either by the purified enzyme or by the crude macrophage cytosol. The peptides, which were resistant to hydrolysis, displayed high affinity for the thimet oligopeptidase as competitive inhibitors. Regardless of the fact that our data do not allow prediction of whether or not a specific peptide would be degraded, it seems very likely that the structural features, which rule out the stability of the MHC class I peptides in the cytosol, may have implications in an optimized repertoire selection for antigen presentation.

Casein pancreatic digest is the basic bacterial growth medium used for diphtheria, botulinum and tetanus toxin vaccine production. It is known that the variation in the peptide content of the casein digest directly affects final toxin yields. In this study, the identification and sequences of eight peptides, four to eight amino acids in length, of casein pancreatic digestion, which seem to be involved in the enhancement of tetanus toxin production, are described. They all contain one or two residues of proline/molecule and a predominance of hydrophobic amino acid residues. The most active peptides show a general structure of Pro-aromatic-Pro, and this pattern resembled the motif displayed by bradykinin-potentiating peptides found in snake venoms. By analogy with the mechanism of bradykinin potentiation through inhibition of the proteolytic degradation of bradykinin, it is suggested that the six peptides identified here could protect the tetanus toxin from proteolysis, once secreted by the bacteria.

A 1.8-kb cDNA clone was isolated from a Bothrops jararaca venom gland cDNA library that encodes a 256-aa precursor for bradykinin-potentiating peptides (angiotensin-converting enzyme inhibitors) and a C-type natriuretic peptide (CNP). The seven bradykinin-potentiating peptides are aligned tandemly after the hydrophobic signal peptide sequence, followed by a putative intervening sequence and a CNP at the C terminus. Northern blot analysis indicated the predominant expression of a 1.8-kb mRNA in the venom glands as well as in the spleen and the brain. Two lower intensity mRNA bands of 3.5 kb and 5.7 kb also hybridized to the cDNA clone. Radioimmunoassay for the CNP was performed using the antiserum against rat CNP. The presence of CNP immunoreactivity was detected in the low molecular weight fraction of the Bothrops jararaca venom.

The heat shock protein [Hsp] family guides several steps during protein synthesis, are abundant in prokaryotic and eukaryotic cells, and are highly conserved during evolution. The Hsp60 family is involved in assembly and transport of proteins, and is expressed at very high levels during autoimmunity or autoinflammatory phenomena. Here, the pathophysiological role of the wild type [WT] and the point mutated K(409)A recombinant Hsp65 of M. leprae in an animal model of Systemic Lupus Erythematosus [SLE] was evaluated in vivo using the genetically homogeneous [NZBxNZW]F(1) mice. Anti-DNA and anti-Hsp65 antibodies responsiveness was individually measured during the animal's life span, and the mean survival time [MST] was determined. The treatment with WT abbreviates the MST in 46%, when compared to non-treated mice [p

Oligopeptidases are tissue endopeptidases that do not attack proteins and are likely to be involved in the maturation and degradation of peptide hormones and neuropeptides. The rabbit brain endooligopeptidase A and the rat testes soluble metallopeptidase (EC 3.4.24.15) are thiol-activated oligopeptidases which are able to generate enkephalin from a number of opiod peptides and to inactivate bradykinin and neurotensin by hydrolyzing the same peptide bonds. A monospecific antibody raised against the purified rabbit brain endooligopeptidase A allowed the identification of a 2.3 kb cDNA coding for a truncated enzyme of 512 amino acids, displaying the same enzymatic features as endooligopeptidase A. In spite of all efforts, employing several strategies, the full-length cDNA could not be cloned until now. The analysis of the deduced amino acid sequence showed no similarity to the rat testes metalloendopeptidase sequence, except for the presence of the typical metalloprotease consensus sequence [HEXXH]. The antibody raised against recombinant endooligopeptidase A specifically inhibited its own activity and reduced the thiol-activated oligopeptidase activity of rabbit brain cytosol to less than 30%. Analysis of the endooligopeptidase A tissue distribution indicated that this enzyme is mainly expressed in the CNS, whereas the soluble metallo EC 3.4.24.15 is mainly expressed in peripheral tissues.