The Experts below are selected from a list of 3372 Experts worldwide ranked by ideXlab platform
Hanslothar Fuchsbauer - One of the best experts on this subject based on the ideXlab platform.
-
the n terminal peptide of the transglutaminase activating Metalloprotease Inhibitor from streptomyces mobaraensis accommodates both inhibition and glutamine cross linking sites
FEBS Journal, 2020Co-Authors: Norbert E Juettner, Stefan Schmelz, Anita Anderl, Felix Colin, Moritz Classen, Felicitas Pfeifer, Andrea Scrima, Hanslothar FuchsbauerAbstract:Streptomyces mobaraensis is a key player for the industrial production of the protein cross-linking enzyme microbial transglutaminase (MTG). Extra-cellular activation of MTG by the transglutaminase-activating Metalloprotease (TAMP) is regulated by the TAMP Inhibitory protein SSTI that belongs to the large Streptomyces subtilisin Inhibitor (SSI) family. Despite decades of SSI research, the binding site for Metalloproteases such as TAMP remained elusive in most of the SSI proteins. Moreover, SSTI is a MTG substrate, and the preferred glutamine residues for SSTI cross-linking are not determined. To address both issues, that is, determination of the TAMP and the MTG glutamine binding sites, SSTI was modified by distinct point mutations as well as elongation or truncation of the N-terminal peptide by six and three residues respectively. Structural integrity of the mutants was verified by the determination of protein melting points and supported by unimpaired subtilisin Inhibitory activity. While exchange of single amino acids could not disrupt decisively the SSTI TAMP interaction, the N-terminally shortened variants clearly indicated the highly conserved Leu40-Tyr41 as binding motif for TAMP. Moreover, enzymatic biotinylation revealed that an adjacent glutamine pair, upstream from Leu40-Tyr41 in the SSTI precursor protein, is the preferred binding site of MTG. This extension peptide disturbs the interaction with TAMP. The structure of SSTI was furthermore determined by X-ray crystallography. While no structural data could be obtained for the N-terminal peptide due to flexibility, the core structure starting from Tyr41 could be determined and analysed, which superposes well with SSI-family proteins. ENZYMES: Chymotrypsin, EC3.4.21.1; griselysin (SGMPII, SgmA), EC3.4.24.27; snapalysin (ScNP), EC3.4.24.77; streptogrisin-A (SGPA), EC3.4.21.80; streptogrisin-B (SGPB), EC3.4.21.81; subtilisin BPN', EC3.4.21.62; transglutaminase, EC2.3.2.13; transglutaminase-activating Metalloprotease (TAMP), EC3.4.-.-; tri-/tetrapeptidyl aminopeptidase, EC3.4.11.-; trypsin, EC3.4.21.4. DATABASES: The atomic coordinates and structure factors (PDB 6I0I) have been deposited in the Protein Data Bank (http://www.rcsb.org).
-
the transglutaminase activating Metalloprotease Inhibitor from streptomyces mobaraensis is a glutamine and lysine donor substrate of the intrinsic transglutaminase
FEBS Letters, 2008Co-Authors: Susan Schmidt, Frank Adolf, Hanslothar FuchsbauerAbstract:Transglutaminase (TGase) from Streptomyces mobaraensis is an extra-cellular enzyme that cross-links proteins to high molecular weight aggregates. Screening for intrinsic substrates now revealed the dual Streptomyces subtilisin Inhibitor-like Inhibitor Streptomyces subtilisin and transglutaminase activating Metalloprotease (TAMEP) Inhibitor (SSTI), equally directed against subtilisin and the TGase activating Metalloprotease TAMEP, is both a glutamine and a lysine donor protein. Reactivity of glutamines is lost during culture, most likely by TGase mediated deamidation, and, accordingly, cross-linking only occurred if SSTI from early cultures was used. Interestingly, release of buried endo-glutamines by the lipoamino acid N-lauroylsarcosine could restore SSTI reactivity. Formation of lipoamino acids by Streptomycetes suggests such compounds could also modulate in vivo TGase mediated SSTI cross-linking.
Roy A Black - One of the best experts on this subject based on the ideXlab platform.
-
TIMP3 checks inflammation
Nature Genetics, 2004Co-Authors: Roy A BlackAbstract:Mice deficient in the Metalloprotease Inhibitor TIMP3, which inhibits the tumor-necrosis factor alpha (TNF-α)-converting enzyme (TACE, also called ADAM17), have elevated levels of TNF and severe inflammation in the liver. This result confirms the physiological importance of the soluble form of TNF and identifies TIMP3 as a crucial regulator of this inflammatory cytokine.
-
tumor necrosis factor alpha converting enzyme tace is a growth hormone binding protein ghbp sheddase the Metalloprotease tace adam 17 is critical for pma induced gh receptor proteolysis and ghbp generation
Endocrinology, 2000Co-Authors: Yue Zhang, Roy A Black, Jing Jiang, Gerhard Baumann, Stuart J. FrankAbstract:The GH binding protein (GHBP), which exists in many vertebrates, is a circulating high affinity binding protein corresponding to the extracellular domain of the GH receptor (GHR). In humans, rabbits, and several other species, the GHBP is generated by proteolysis of the GHR and shedding of its extracellular domain. We previously showed that GHBP shedding is inducible by the phorbol ester phorbol 12-myristate,13-acetate (PMA) and inhibited by the Metalloprotease Inhibitor, Immunex Corp. Compound 3 (IC3). The metzincin Metalloprotease, tumor necrosis factor-α (TNF-α)-converting enzyme (TACE), catalyzes the shedding of TNF-α from its transmembrane precursor, a process that is also inhibitable by IC3. TACE may hence be a candidate for GHBP sheddase. In this study, we reconstitute fibroblasts derived from a TACE knockout mouse (Null cells) with either the rabbit (rb) GHR alone (Null/R) or rbGHR plus murine TACE (Null/R+T). Although GHR in both cells was expressed at similar abundance, dimerized normally and ca...
-
Blockade of growth hormone receptor shedding by a Metalloprotease Inhibitor.
Endocrinology, 1998Co-Authors: Jimmy Alele, Roy A Black, Jing Jiang, Jeffrey F. Goldsmith, Xiaoyong Yang, Hiralal G. Maheshwari, Gerhard Baumann, Stuart J. FrankAbstract:GH, an important growth-promoting and metabolic hormone, exerts its biological effects by interacting with cell surface GH receptors (GHRs). The GHR is a single membrane-spanning protein that binds GH via its extracellular domain. The high affinity GH-binding protein (GHBP), which corresponds to a soluble form of the GHR extracellular domain, carries a substantial fraction of the GH in the circulation of various species and probably has a role in modulation of the hormone’s bioavailability. Although in rodents, it is believed that the GHBP is largely derived by translation of an alternatively spliced GHR messenger RNA, in humans and rabbits, proteolytic cleavage of the membrane-anchored receptor releases the GHR extracellular domain, which is believed to thereby become the GHBP. In this study, we used human IM-9 lymphocytes and GHR antibodies to study this proteolytic shedding of the GHBP. As determined by immunoblotting with anti-GHR cytoplasmic domain serum, addition of phorbol 12-myristate 13-acetate (...
-
A Metalloprotease Inhibitor blocks shedding of the IL-6 receptor and the p60 TNF receptor.
Journal of immunology (Baltimore Md. : 1950), 1995Co-Authors: Jürgen Müllberg, Roy A Black, F H Durie, Carol Otten-evans, Mark Alderson, Stefan Rose-john, David Cosman, Kendall M. MohlerAbstract:Many cytokines and soluble cytokine receptors are generated by limited proteolysis of membrane-bound precursors. We have examined the ability of the recently described Metalloprotease Inhibitor, TNF-alpha protease Inhibitor (TAPI), and other protease Inhibitors to modulate shedding. The membrane-bound forms of the ligands TNF-alpha and CSF-1, the p60 TNFR and the IL-6R, were expressed in COS-7 cells. As expected, TAPI blocked the spontaneous and PMA-induced release of TNF-alpha from transfected cells. Interestingly, TAPI also inhibited the release of soluble forms of p60 TNFR and IL-6R in COS-7 cells. However, the processing of CSF-1, which also requires proteolytic cleavage of a membrane protein, was not affected. The ability of TAPI to inhibit shedding was unique, since several other classes of protease Inhibitors, including three other Metalloprotease Inhibitors, did not inhibit shedding of IL-6R. To determine whether TAPI would prevent shedding under more physiologic conditions, we demonstrated that TAPI was able to prevent unstimulated and PMA-induced release of the soluble forms of TNF-alpha, p60 TNFR, and IL-6R from the monocytic cell line, THP-1, and from human peripheral blood monocytes. In addition, TAPI was able to inhibit LPS-induced shedding of the p60 TNFR and TNF-alpha from monocytes. In summary, our results indicate that a Metalloprotease or group of related Metalloproteases is responsible for the proteolytic cleavage of several cell surface proteins.
-
a Metalloprotease Inhibitor blocks shedding of the 80 kd tnf receptor and tnf processing in t lymphocytes
Journal of Experimental Medicine, 1995Co-Authors: Paul D Crowe, Roy A Black, Kendall M. Mohler, Barbara N Walter, Carol Ottenevans, Carl F WareAbstract:TNF is synthesized as a 26-kD membrane-anchored precursor and is proteolytically processed at the cell surface to yield the mature secreted 17-kD polypeptide. The 80-kD tumor necrosis factor (TNF) receptor (TNFR80) is also proteolytically cleaved at the cell surface (shed), releasing a soluble ligand-binding receptor fragment. Since processing of TNF and TNFR80 occurs concurrently in activated T cells, we asked whether a common protease may be involved. Here, we present evidence that a recently described Inhibitor of TNF processing N-(D,L-[2-(hydroxyaminocarbonyl)methyl]-4-methylpentanoyl)L- 3-(2'naphthyl)- alanyl-L-alanine, 2-aminoethyl amide (TAPI) also blocks shedding of TNFR80, suggesting that these processes may be coordinately regulated during T cell activation. In addition, studies of murine fibroblasts transfected with human TNFR80, or a cytoplasmic deletion form of TNFR80, reveal that inhibition of TNFR80 shedding by TAPI is independent of receptor phosphorylation and does not require the receptor cytoplasmic domain.
Andrea C Leblanc - One of the best experts on this subject based on the ideXlab platform.
-
effect of tumor necrosis factor alpha converting enzyme tace and Metalloprotease Inhibitor on amyloid precursor protein metabolism in human neurons
Journal of Neurochemistry, 2002Co-Authors: Megan Blacker, Mark C Noe, Thomas J Carty, Cynthia G Goodyer, Andrea C LeblancAbstract:Tumor necrosis factor-alpha (TNF-alpha) is implicated in inflammatory processes and much effort is being directed at inhibiting the release of TNF-alpha for treatment of inflammatory conditions. In this context, the drug CP-661,631 has been developed to inhibit the TNF-alpha converting enzyme (TACE). However, TACE is also implicated in amyloid precursor protein secretion. Amyloid precursor protein (APP) undergoes constitutive and regulated secretion by alpha-secretase endoproteolytic cleavage within the amyloid beta peptide (Abeta) domain. Alternative cleavage at the N- and C-terminus of the Abeta domain by beta- and gamma-secretases results in the production of Abeta. In many cellular and in vivo animal models, increased secretion of APP results in a concomitant decrease in the production of Abeta suggesting that the two pathways are intricately linked. However, in human primary neuron cultures, increased APP secretion is not associated with a decrease in total Abeta production. To determine if the use of CP-661,631 may enhance amyloidogenic processing in human brain, we have assessed the effect of CP-661,631 on APP metabolism in primary cultures of human neurons. Our results show that CP-661,631 effectively prevents regulated APP secretion but does not increase total Abeta levels in human primary neuron cultures.
Charles B Shoemaker - One of the best experts on this subject based on the ideXlab platform.
-
small molecule Metalloprotease Inhibitor with in vitro ex vivo and in vivo efficacy against botulinum neurotoxin serotype a
Toxicon, 2017Co-Authors: Alan R Jacobson, Michael Adler, Nicholas R Silvaggi, Karen N Allen, Genessa M Smith, Ross A Fredenburg, Ross L Stein, Jongbeak Park, Xiaochuan Feng, Charles B ShoemakerAbstract:Botulinum neurotoxins (BoNTs) are the most toxic substances known to mankind and are the causative agents of the neuroparalytic disease botulism. Their ease of production and extreme toxicity have caused these neurotoxins to be classified as Tier 1 bioterrorist threat agents and have led to a sustained effort to develop countermeasures to treat intoxication in case of a bioterrorist attack. While timely administration of an approved antitoxin is effective in reducing the severity of botulism, reversing intoxication requires different strategies. In the present study, we evaluated ABS 252 and other mercaptoacetamide small molecule active-site Inhibitors of BoNT/A light chain using an integrated multi-assay approach. ABS 252 showed Inhibitory activity in enzymatic, cell-based and muscle activity assays, and importantly, produced a marked delay in time-to-death in mice. The results suggest that a multi-assay approach is an effective strategy for discovery of potential BoNT therapeutic candidates.
Susan Schmidt - One of the best experts on this subject based on the ideXlab platform.
-
the transglutaminase activating Metalloprotease Inhibitor from streptomyces mobaraensis is a glutamine and lysine donor substrate of the intrinsic transglutaminase
FEBS Letters, 2008Co-Authors: Susan Schmidt, Frank Adolf, Hanslothar FuchsbauerAbstract:Transglutaminase (TGase) from Streptomyces mobaraensis is an extra-cellular enzyme that cross-links proteins to high molecular weight aggregates. Screening for intrinsic substrates now revealed the dual Streptomyces subtilisin Inhibitor-like Inhibitor Streptomyces subtilisin and transglutaminase activating Metalloprotease (TAMEP) Inhibitor (SSTI), equally directed against subtilisin and the TGase activating Metalloprotease TAMEP, is both a glutamine and a lysine donor protein. Reactivity of glutamines is lost during culture, most likely by TGase mediated deamidation, and, accordingly, cross-linking only occurred if SSTI from early cultures was used. Interestingly, release of buried endo-glutamines by the lipoamino acid N-lauroylsarcosine could restore SSTI reactivity. Formation of lipoamino acids by Streptomycetes suggests such compounds could also modulate in vivo TGase mediated SSTI cross-linking.