Katayama S, Betheil J J, Seifter S
Biochim Biophys Acta. 1978 May 11;524(1):188-97. doi: 10.1016/0005-2744(78)90117-1.
Purified collagenase of Clostridium histolyticum was shown to cleave reduced and S-carboxamidomethylated bovine neurophysin between Cys-13 and Gly-14. The scission resulted in formation of two separable fragments: a smaller peptide arising from residues 1 through 13, and a larger peptide comprising the remainder of the residues of the protein. By dansylation procedures, the smaller peptide was shown to have amino-terminal alanine as expected from the sequence of neurophysin II, and the larger peptide had amino-terminal glycine as anticipated. These results show that collagenase indeed cleaves bovine neurophysin II in accord with the specificity postulated for that enzyme, i.e., scission between -X-Gly- in a sequence of -Pro-X-Gly-Pro-Y-. This result, obtained with a non-collagenous protein substrate, is further confirmation of the specificity of collagenase as established by its action on collagens and on synthetic oligopeptides.
溶组织梭菌纯化胶原酶可在半胱氨酸 -13和甘氨酸 -14之间切割还原型和 S-羧甲基化牛神经垂体素。这种切割导致形成两个可分离的片段：一个较小的肽段由 1至 13位残基组成，一个较大的肽段包含该蛋白质其余的残基。通过丹磺酰化程序，较小的肽段显示其氨基末端为丙氨酸，这与神经垂体素 II的序列预期一致，而较大的肽段氨基末端为甘氨酸，正如预期的那样。这些结果表明，胶原酶确实按照该酶假定的特异性切割牛神经垂体素 II，即在 -Pro-X-Gly-Pro-Y-序列中的 -X-Gly-之间进行切割。用非胶原蛋白质底物获得的这一结果，进一步证实了胶原酶通过其对胶原蛋白和合成寡肽的作用所确定的特异性。
