Matrix metalloproteases are secreted by mammalian cells as zymogens and, upon activation, initiate tissue remodeling proteolytic degradation of collagens and proteoglycans. Activation the proenzymes interaction with their specific inhibitors determine net enzymatic activity in extracellular space. We have previously demonstrated that 72T4Cl can be activated a plasma membrane-dependent mechanism for this enzyme. Here, we report purification membrane activator 72T4Cl, which is new...

We have reported that SV40-transformed human lung fibroblasts secrete a 92-kDa metalloprotease which is not detectable in the parental cell line IMR-90. now present complete structure of this enzyme along with evidence it identical to secreted by normal alveolar macrophages, phorbol ester-differentiated monocytic leukemia U937 cells, fibrosarcoma HT1080 and cultured keratinocytes. A similar, perhaps identical, can be released polymorphonuclear cells. The preproenzyme synthesized as...

H-ras-transformed human bronchial epithelial cells (TBE-1) secrete a single major extracellular matrix metalloprotease which is not found in the normal parental cells. The enzyme secreted latent form of 72 kDa, can be activated to catalyze cleavage basement membrane macromolecule type IV collagen. substrates their order preference are: gelatin, collagen, V fibronectin, and VII collagen; but does cleave interstitial collagens or laminin. This protease identical gelatinase isolated from skin...

The preferred antitubercular drug isoniazid specifically targets a long-chain enoyl-acyl carrier protein reductase (InhA), an enzyme essential for mycolic acid biosynthesis in Mycobacterium tuberculosis. Despite the widespread use of this more than 40 years, its precise mode action has remained obscure. Data from x-ray crystallography and mass spectrometry reveal that mechanism against InhA is covalent attachment activated form to nicotinamide ring adenine dinucleotide bound within active site InhA.

Simian virus 40 (SV40)-transformed human lung fibroblasts secrete both 72-kDa type IV collagenase and a closely related 92-kDa that was not detected in the parental cell line. The procollagenase purified from these cells exists noncovalent complex with tissue inhibitor of metalloproteases, TIMP. Here we report HRAS-transformed bronchial epithelial cells, SV40-transformed fibroblasts, normal skin stable but stoichiometric 24-kDa referred to here as "TIMP-2." TIMP-2 is TIMP, demonstrated by...

We present a cascade of proteolytic events catalyzed by the proteases secreted cultured keratinocytes and fibroblasts that results in activation interstitial procollagenase. Cultured human skin constitutively secrete collagenase stromelysin as proenzymes. In contrast, found serum-free organ culture conditioned medium is activated. Cocultivation major cellular components culture, dermal epidermal keratinocytes, induces procollagenase prostromelysin presence plasminogen. This occurs through...

We have purified and determined the complete primary structure of human stromelysin, a secreted metalloprotease with wide range substrate specificities. Human stromelysin is synthesized in preproenzyme form calculated size 53,977 Da 17-amino acid long signal peptide. Prostromelysin two forms, apparent molecular masses on NaDodSO4/PAGE 60 57 kDa. The minor 60-kDa polypeptide glycosylated major 57-kDa protein containing N-linked complex oligosaccharides. Zymogen activation by trypsin results...

A monoclonal antibody (C6.7) has been generated against the calcium-replete form of human platelet thrombospondin (TSP). C6.7 is specific for TSP as determined by both competitive radioimmunoassay and immunoprecipitation. This inhibits thrombin- A23187-induced aggregation gel-filtered platelets in a concentration-dependent manner without affecting secretion serotonin. The epitope on recognized localized to an 18-kDa fragment that present mild chymotryptic digests TSP. disulfide-linked 120-...

The effects of diabetes on collagen cross-link formation and solubility were investigated in granulation tissue induced by polyester fabric implanted subcutaneously rats at the same time was produced injection streptozotocin. Thus, all analyzed formed a diabetic milieu. Ten days later implants removed total content as well fraction soluble 0.5 M acetic acid determined. Predominantly type I accumulated implants. Total diabetics controls; however, acid-soluble animals only half that controls...

Type IV collagenase (gelatinase) readily cleaves denatured collagen into very small peptides. Large cyanogen bromide fragments (25 kDa) of type I are degraded at the same rate as complete alpha-chain. A number gelatinolytic cleavage sites alpha 1(I)CB7 and 1(I)CB8, representing 50% alpha-chain, were determined by sequence analysis product In addition to expected between glycine hydrophobic residues, several other identified. These Gly-Glu, Gly-Asn, Gly-Ser. Basic residues found adjacent site...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXT.kappa.-Bungarotoxin: complete amino acid sequence of a neuronal nicotinic receptor probeGregory A. Grant and Vincent ChiappinelliCite this: Biochemistry 1985, 24, 6, 1532–1537Publication Date (Print):March 12, 1985Publication History Published online1 May 2002Published inissue 12 March 1985https://pubs.acs.org/doi/10.1021/bi00327a036https://doi.org/10.1021/bi00327a036research-articleACS PublicationsRequest reuse permissionsArticle...

The nucleotide sequence of serA, the structural gene Escherichia coli which codes for D-3-phosphoglycerate dehydrogenase, has been determined. contains 1233 nucleotides code 409 amino acids subunit tetrameric enzyme, as well initiator methionine, is cleaved from mature protein, and termination codon. majority primary structure enzyme confirmed by automated Edman degradation peptide fragments produced a variety cleavage agents. Comparison acid phosphoglycerate dehydrogenase with other...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTAmino acid sequence of a collagenolytic protease from the hepatopancreas fiddler crab, Uca pugilatorGregory A. Grant, Kenneth O. Henderson, Arthur Z. Eisen, and Ralph BradshawCite this: Biochemistry 1980, 19, 20, 4653–4659Publication Date (Print):September 30, 1980Publication History Published online1 May 2002Published inissue 30 September 1980https://pubs.acs.org/doi/10.1021/bi00561a018https://doi.org/10.1021/bi00561a018research-articleACS...

Thrombospondin (TSP) cDNA clones were isolated from a human fibroblast library by screening with degenerate synthetic oligodeoxynucleotides. The probes designed based on the sequences of peptides tryptic digests TSP. largest clone identified contains an open reading frame that encodes amino-terminal heparin binding domain TSP and part immediately adjacent collagen domain, region includes site disulfide crosslinking responsible for trimer formation. In addition to known sequence mature (which...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTPurification of an apparent rat liver prothrombin precursor. Characterization and comparison to normal prothrombinCharles T. Esmon, Gregory A. Grant, John W. SuttieCite this: Biochemistry 1975, 14, 8, 1595–1600Publication Date (Print):April 22, 1975Publication History Published online1 May 2002Published inissue 22 April 1975https://pubs.acs.org/doi/10.1021/bi00679a008https://doi.org/10.1021/bi00679a008research-articleACS PublicationsRequest reuse...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTA collagenolytic serine protease with trypsin-like specificity from the fiddler crab Uca pugilatorGregory A. Grant, James C. Sacchettini, and Howard G. WelgusCite this: Biochemistry 1983, 22, 2, 354–358Publication Date (Print):January 18, 1983Publication History Published online1 May 2002Published inissue 18 January 1983https://pubs.acs.org/doi/10.1021/bi00271a019https://doi.org/10.1021/bi00271a019research-articleACS PublicationsRequest reuse...