Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.24.23 (MMP)
 4,246 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Recombinant human fibroblast pro-MMP-3 (prostromelysin-1) expressed in Chinese hamster ovary cells and the zymogen from cultured human dermal fibroblasts have been purified by monoclonal antibody immunoaffinity chromatography, and the role of Ca2+ in proenzyme activation and thermostability of the low mass catalytic domain of MMP-3 has been investigated. In the presence of high Ca2+ (5.0 mM), the organomercurial aminophenylmercuric acetate (APMA) initiated the stepwise removal of both NH2- and COOH-terminal domains from both recombinant and dermal fibroblast proenzymes, resulting in the generation of a heterogeneous family of nonglycosylated low mass truncated active enzyme species beginning at Phe83. However, in the presence of low Ca2+ (0.1 mM), incubation of recombinant pro-MMP-3 with or without APMA did not result in formation of either the high or low mass forms of active MMP-3 but resulted in complete autolysis of both enzyme species. The concentration of Ca2+ required for optimal pro-MMP-3 activation and stability of the low mass catalytic domain was 2.0 mM. The low mass truncated enzyme species containing the catalytic domain were remarkably heat-stable (90 min at 60 degrees C) in high Ca2+ (5.0 mM) but rapidly autolyzed when heated at 60 degrees C in low Ca2+ (0.1 mM). The thermostability properties of MMP-3 appeared to be specific for Ca2+, since no other divalent metal ions tested were able to confer thermostability to the low mass catalytic domain of MMP-3. From homology to the thermostable bacterial metalloprotease, thermolysin, two putative Ca2+ binding sites were found in the catalytic domain of MMP-3 and several other members of the MMP gene family. These putative Ca2+ binding sites are postulated to play an important role in stabilizing active MMP-3 and other members of the matrix metalloprotease gene family by protecting them against autolysis.
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PMID:Recombinant Chinese hamster ovary cell matrix metalloprotease-3 (MMP-3, stromelysin-1). Role of calcium in promatrix metalloprotease-3 (pro-MMP-3, prostromelysin-1) activation and thermostability of the low mass catalytic domain of MMP-3. 844 Jul 30

Cultured equine lamellar hoof explants secrete the pro-enzymes matrix metalloproteinase-2 (MMP-2, 72 kDa) and MMP-2 (92 kDa). Untreated explants remained intact when tested on a calibrated force transducer, but when treated with an MMP activator, developed "in-vitro laminitis", separating at the dermal-epidermal junction. Explants treated with the bacterial protease thermolysin separated dose-dependently; this was accompanied by activation of both MMP-2 and -9. Thermolysin-mediated MP activation did not occur in a cell-free system and was not inhibited by the addition of the MMP inhibitor and batimastat. These findings suggest that thermolysin-mediated gelatinase activation is not dependent on membrane-bound matrix metalloproteinase (MT-MMP) activation, providing further evidence that bacteria can produce potent MMP activators that probably facilitate host invasion.
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PMID:Thermolysin activates equine lamellar hoof matrix metalloproteinases. 1181 17

Streptomyces caespitosus neutral protease (ScNP) is one of the smallest metalloproteinase with a molecular mass of 14 kDa. Effects of solvent composition on ScNP activity were examined using a peptide substrate. The k(cat)/K(m) values of ScNP exhibited bell-shaped pH-dependence with the optimal pH of 6.4-7.0 and the pK(a) values of 5.0 +/- 0.1 and 8.3 +/- 0.1. ScNP activity increased in an exponential fashion with increasing [NaCl]. The relative k(cat)/K(m) value at 3.6 M NaCl to that at 0 M NaCl was 3.7, and the degree of the activation at x M NaCl was expressed as 1.2 (x) (x < 2.0) and 1.4(x) (x > 2.0). On the other hand, ScNP activity decreased with increasing concentrations of LiCl, KCl, NaBr, LiBr, KBr and NaClO(4). Alcohols inhibited ScNP activity with the IC(50) values, the concentration required for decreasing the activity at 50% of the maximum, of 0.77-6.54 M. The order of the inhibitory potency was 1-butanol, 2-methyl-1-propanol, 2-methyl-2-butanol > 2-methyl-2-propanol, 2-butanol, 1-propanol > 2-propanol >> ethanol >> methanol. The activities recovered completely by the dilution of alcohols, suggesting that the ScNP inhibition by alcohols is reversible. These characteristics of ScNP are compared with those of human matrix metalloproteinase 7 and thermolysin.
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PMID:Effects of neutral salts and alcohols on the activity of Streptomyces caespitosus neutral protease. 1764 80