Trocheris I, Herry P, Keil-Dlouha V, Keil B
Biochim Biophys Acta. 1980 Oct;615(2):436-48. doi: 10.1016/0005-2744(80)90510-0.
A study of the influence of chemical modifications on the activity of Achromobacter iophagus collagenase (EC 3.4.24.8) has led to the following conclusions: a modification of 4 out of 80 COOH groups with carbodiimide led to 90% loss of enzymic activity. A 70% inactivation was found after modification of two tyrosines out of 30 with tetranitromethane. The modification of four to six tryptophans out of 16 with 2-hydroxy-5-nitrobenzyl bromide decreased enzyme activity to 36%. This inactivation is accelerated in the presence of collagen. An increase of reagent/enzyme molar ratio led to a modification of 16 tryptophan residues and denaturation of Acahromobacter collagenase. A modification of two arginines out of 18 with 1,2-cyclohexanedione and eight NH2 groups out of 24 with 2,3-dimethyl maleic anhydride does not change the collagenolytic activity. All NH2 groups become available for 2,3-dimethyl maleic anhydride after dissociation of the dimer. A possible analogy of hydrolytic site of collagenase with that of two other known bacterial metalloproteinases (thermolysin and Bacillus subtilis neutral proteinase (EC 3.4.24.4)) is discussed.
一项关于化学修饰对食油无色杆菌胶原酶（EC 3.4.24.8）活性影响的研究得出了以下结论：用碳二亚胺对80个COOH基团中的4个进行修饰导致酶活性丧失90%。用四硝基甲烷对30个酪氨酸中的2个进行修饰后，发现失活70%。用2-羟基-5-硝基苄基溴对16个色氨酸中的4至6个进行修饰，使酶活性降至36%。在胶原蛋白存在的情况下，这种失活会加速。试剂/酶摩尔比的增加导致16个色氨酸残基被修饰，食油无色杆菌胶原酶变性。用1,2-环己二酮对18个精氨酸中的2个进行修饰，用2,3-二甲基马来酸酐对24个NH₂基团中的8个进行修饰，不会改变胶原olytic活性。二聚体解离后，所有NH₂基团都可与2,3-二甲基马来酸酐反应。文中讨论了胶原酶水解位点与另外两种已知细菌金属蛋白酶（嗜热菌蛋白酶和枯草芽孢杆菌中性蛋白酶（EC 3.4.24.4））水解位点可能的相似性。
