Kajiwara K, Fujita A, Tsuyuki H, Kumazaki T, Ishii S
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Hokkaido University.
J Biochem. 1991 Sep;110(3):350-4. doi: 10.1093/oxfordjournals.jbchem.a123584.
Streptomyces griseus metalloendopeptidase II (SGMPII) was shown to form tight complexes with several Streptomyces protein inhibitors which had been believed to be specific to serine proteases, such as Streptomyces subtilisin inhibitor (SSI), plasminostreptin (PS), and alkaline protease inhibitor-2c' (API-2c'), as well as with Streptomyces metalloprotease inhibitor (SMPI). The dissociation constants of complexes between SGMPII and these inhibitors were successfully determined by using a novel fluorogenic bimane-peptide substrate. The values ranged from nM to pM. The results of studies by gel chromatographic and enzymatic analyses indicated that SGMPII is liberated from the complex with SSI by the addition of subtilisin BPN'. SGMPII and subtilisin BPN' proved, therefore, to interact with SSI in a competitive manner, despite the difference in the chemical nature of their active sites.
灰色链霉菌金属内肽酶II（SGMPII）已被证明能与几种链霉菌蛋白抑制剂形成紧密复合物，这些抑制剂曾被认为是丝氨酸蛋白酶特异性抑制剂，如枯草芽孢杆菌链霉菌抑制剂（SSI）、纤溶酶链菌素（PS）和碱性蛋白酶抑制剂-2c'（API-2c'），以及链霉菌金属蛋白酶抑制剂（SMPI）。通过使用一种新型的荧光双硫腙肽底物，成功测定了SGMPII与这些抑制剂之间复合物的解离常数。其值范围从纳摩尔到皮摩尔。凝胶色谱和酶分析研究结果表明，通过添加枯草杆菌蛋白酶BPN'，SGMPII可从与SSI的复合物中释放出来。因此，尽管SGMPII和枯草杆菌蛋白酶BPN'的活性位点化学性质不同，但它们与SSI以竞争方式相互作用。
