Pivnenko T N, Epshteĭn L M, Kolodzeĭskaia M V, Kudinov S A
Prikl Biokhim Mikrobiol. 1989 Jul-Aug;25(4):490-7.
Trypsin from pyloric caeca of Pacific salmon was purified by affinity chromatography of the water extract on hexamethylenediamine-glycidylmethacrylate-cellulose. A protein band with a molecular weight of 22.5 kDa was found on SDS-electrophoresis in PAG. The protein band was homogeneous according to isoelectrofocusing in PAG (pI 4.0). The amino acid composition of the enzyme is typical of trypsin anionic forms; the major difference from the cationic forms is the lower content of lysine. The differences in properties caused by change of the enzyme molecule charge are similar to those observed in cationic trypsin when the lysine epsilon-amino groups of the latter are modified (change of pI, shift of the pH-optimum towards basic values, increase of stability to autolysis). Some natural trypsin inhibitors of the different origin suppressed the enzyme activity of trypsin from Pacific salmon in typical stoichiometric ratios. An unusual interaction of the enzyme with the specific inhibitor N-L-tosyl-L-lysine chloromethyl ketone was observed.
采用己二胺-甲基丙烯酸缩水甘油酯-纤维素对太平洋鲑鱼幽门盲囊中的胰蛋白酶进行水提取物亲和层析纯化。在聚丙烯酰胺凝胶（PAG）的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-电泳）上发现一条分子量为22.5 kDa的蛋白条带。根据在PAG中的等电聚焦（pI 4.0），该蛋白条带是均一的。该酶的氨基酸组成是典型的阴离子型胰蛋白酶；与阳离子型的主要区别在于赖氨酸含量较低。酶分子电荷变化引起的性质差异与阳离子型胰蛋白酶赖氨酸ε-氨基被修饰时观察到的差异相似（pI变化、pH最适值向碱性值偏移、对自溶稳定性增加）。不同来源的一些天然胰蛋白酶抑制剂以典型的化学计量比抑制太平洋鲑鱼胰蛋白酶的酶活性。观察到该酶与特异性抑制剂N-L-甲苯磺酰-L-赖氨酸氯甲基酮之间存在异常相互作用。
