3.4.24.27 Ba2+ activates 8% at 1 mM, 15% at 5 mM 734335 3.4.24.27 Ca2+ - 683276, 683691 3.4.24.27 Ca2+ 4 calcium ions required for structural stability 651799 3.4.24.27 Ca2+ activates 36% at 1 mM, 48% at 5 mM 734335 3.4.24.27 Ca2+ CaCl2 does not exert any significant influence on the activity of immobilized thermolysin, but stabilizes the enzyme in absence of Zn2+ 733524 3.4.24.27 Ca2+ contains 4 structural calcium ions 649825 3.4.24.27 Ca2+ four Ca2+ ions are required for thermostability 711460 3.4.24.27 Ca2+ four Ca2+ per enzyme molecule are required for enzyme stability 683276 3.4.24.27 Ca2+ four ions per enzyme molecule required for stability 683182, 683615, 683616, 683687, 684019 3.4.24.27 Ca2+ protease binds calcium ions in the regions that are involved in the autolytic maturation process, at least one of the calcium ions plays a regulatory role. This calcium ion plays an important role as a switch that modulates the protease between stable and unstable states as appropriate to the biological need 718367 3.4.24.27 Ca2+ required 717283, 717396 3.4.24.27 Ca2+ required for stability 683190 3.4.24.27 Ca2+ stablizes the enzyme 734109 3.4.24.27 Ca2+ there are two adjacent calcium ions seemingly firmly bound inside the surface of the molecule by chelation to five acidic groups: Asp138, Glu177, Asp185, Glu190 and Asp191, two additional calcium binding sites are at exposed surface regions, one chelated by Asp57 and possibly also by Asp59 and the other chelated by Asp200 31146 3.4.24.27 Ca2+ thermolysin consists of four Ca2+ ligand ions necessary for stability 712856 3.4.24.27 Ca2+ thermolysin has four calcium ions responsible for its thermostability 711724 3.4.24.27 Ca2+ TLN contains one zinc ion and four calcium ions, and these ions contribute to enzymatic catalysis and structural stability 717113 3.4.24.27 CaCl2 required 734411 3.4.24.27 Co2+ 2fold activation at 0.4 mM 650286 3.4.24.27 Co2+ active zinc ion in thermolysin can be substituted by Co2+, doubles activity 651789 3.4.24.27 Co2+ increases the activity 3-4fold at up to 2 mM, but inhibits at 2-18 mM, activation-and-inhibition dual effects of Co2+ ion are analysed kinetically, Co2+-dependent activation is inhibited competitively by Zn2+ ion at 0.0001-0.001 mM 683615 3.4.24.27 Co2+ the active site zinc atom of thermolysin is replaced by cobalt 717283 3.4.24.27 K+ activates, preference of monovalent cations in descending order: Na+, K+, Li+ 683614 3.4.24.27 Li+ activates, preference of monovalent cations in descending order: Na+, K+, Li+ 683614 3.4.24.27 Mg2+ activates 12% at 1 mM, 37% at 5 mM 734335 3.4.24.27 Mn2+ activates 12% at 1 mM, 25% at 5 mM 734335 3.4.24.27 additional information mechanism of salt-induced activation 683614 3.4.24.27 additional information thermolysin activity as well as its stability is remarkably enhanced by high concentration of neutral salts consisting of Na+, K+, Cl-, and Br- in the synthesis and hydrolysis of N-carbobenzoxy-L-aspertyl-L-phenylalanine methyl ester and hydrolysis of N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide 683615 3.4.24.27 Na+ 4 M Na+ stimulates the hydrolytic activity of wild type thermolysin about 13fold 712532 3.4.24.27 Na+ activates, preference of monovalent cations in descending order: Na+, K+, Li+, the bell-shaped pH dependence profile of the FAGLA-hydrolyzing activity of thermolysin is shifted from pH 5.4 to pH 6.7 by the addition of 4 M NaCl 683614 3.4.24.27 NaBr 4 M enhances activity 13-15 times 651789 3.4.24.27 NaCl 4 M enhances activity 13-15 times 651789 3.4.24.27 NaCl activates 683182 3.4.24.27 NaCl activates at 3 M 733524 3.4.24.27 NaCl activates the mutant enzymes at 4 M to 17-19fold of wild-type enzyme activity, overview 683190 3.4.24.27 NaCl activity is remarkably enhanced by 1-5 M neutral salts 651745 3.4.24.27 NaCl induces activation 651799 3.4.24.27 NaCl induces enzyme activation, activation of mutant enzymes is reduced compared to the wild-type enzyme 717396 3.4.24.27 NaCl required 734411 3.4.24.27 NaSCN 3fold increase in catalytic activity of thermolysin when the NaSCN concentration is increased to 1 M, but decrease in catalytic activity at higher concentrations of NaSCN 717283 3.4.24.27 Zinc the three zinc ligands are two histidines and glutamic acid 31146 3.4.24.27 Zinc zinc metalloproteinase 31137, 31141, 31142 3.4.24.27 Zn2+ - 683691 3.4.24.27 Zn2+ a single catalytic Zn2+ ion is essential for hydrolytic activity 711460 3.4.24.27 Zn2+ a zinc metalloprotease 683701, 733524, 733899 3.4.24.27 Zn2+ a zinc metalloproteinase 683276 3.4.24.27 Zn2+ a zinc metalloproteinase that contains a HEXXH motif, one Zn2+ per enzyme molecule is required for activity 683276 3.4.24.27 Zn2+ a Zn2+ metalloprotease 732993 3.4.24.27 Zn2+ active zinc ion in thermolysin 651789 3.4.24.27 Zn2+ comparison of metal preferences of Escherichia coli peptide deformylase and Bacillus thermoproteolyticus thermolysin. Both enzymes catalyze via the same chemical steps, and reproduce their different preferences for zinc or iron as competent cofactors. In thermolysin, the substrate is strongly activated and can serve as the fifth coordination ligand of zinc prior to the chemical steps. When iron replaces zinc, its stronger interaction with the hydroxide ligand may lead to higher activation barrier in thermolysin 699764 3.4.24.27 Zn2+ contains Zn2+ 711724 3.4.24.27 Zn2+ dependent on 711393 3.4.24.27 Zn2+ dependent on, thermolysin is a bacterial zinc metalloproteinase. The active site zinc atom is tetrahedrally coordinated when the inhibitors N-benzyloxycarbonyl-tryptophan or N-benzyloxycarbonyl-phenylalanine are bound to thermolysin 717283 3.4.24.27 Zn2+ metalloprotease 717283 3.4.24.27 Zn2+ one ion per enzyme molecule essential for activity, the enzyme contains the HEXXH motif constituting the zinc catalytic site 683687 3.4.24.27 Zn2+ one ion per enzyme molecule required for activity 683182, 683615, 683616 3.4.24.27 Zn2+ required for activity 683190 3.4.24.27 Zn2+ required for enzyme activity 651799 3.4.24.27 Zn2+ residues H142, H146, and E166 coordinate the catalytic zinc 699468 3.4.24.27 Zn2+ stereochemical relationships between Gln128, Glu143, Gln225, Asp226, His231 and active site Zn2+ of thermolysin, overview 683614 3.4.24.27 Zn2+ the enzyme has a catalytic zinc ion at the active site cleft with a tetrahedral coordination formed by the two histidines of a HEXXH motif, and a glutamic acid located 18-72 residues C-terminal of the HEXXH motif. The fourth zinc coordinating ligand in the free enzyme is a water molecule 733787 3.4.24.27 Zn2+ thermolysin consists of three Zn2+ ligand ions necessary for activity 712856 3.4.24.27 Zn2+ TLN contains one zinc ion and four calcium ions, and these ions contribute to enzymatic catalysis and structural stability 717113 3.4.24.27 Zn2+ zinc endopeptidase 653830 3.4.24.27 Zn2+ zinc metalloprotease, no efect on activity by exogenous Zn2+ at 1 mM, inhibitory at 5 mM 734335 3.4.24.27 Zn2+ zinc metalloproteinase, one ion per enzyme molecule required for activity 684019 3.4.24.27 Zn2+ zinc-containing neutral metalloendoprotease 650540 3.4.24.27 Zn2+ zinc-metallopeptidase 649825 3.4.24.27 Zn2+ Zn-dependent 650286