Dahlmann B, Rutschmann M, Kuehn L, Reinauer H
Biochem J. 1985 May 15;228(1):171-7. doi: 10.1042/bj2280171.
A multicatalytic proteinase from rat skeletal muscle contains active site(s) catalysing the degradation of benzoyl-Val-Gly-Arg 4-methyl-7-coumarylamide, succinyl-Ala-Ala-Phe 4-methylcoumarylamide and [14C]methylcasein as well as benzyloxy-carbonyl-Leu-Leu-Glu 2-naphthylamide. These activities are 7-14-fold activated by 1 mM-sodium dodecyl sulphate. The activation leads to a higher susceptibility to the proteinase inhibitor chymostatin and to a lower ability to be inhibited and precipitated by antibodies raised against the non-activated enzyme. Since no changes in Mr or subunit composition were observed in the SDS-activated form, some conformational changes seem to occur during the activation step. More pronounced activation was observed in the presence of physiological concentrations of fatty acids; oleic acid at 100 microM concentrations stimulated the proteinase about 50-fold. In contrast with the non-activated proteinase, the activated enzyme considerably degrades muscle cytoplasmic proteins in vitro. Thus it is not unlikely that, in vivo, potential activators such as fatty acids can induce the multicatalytic proteinase to participate in muscle protein breakdown.
大鼠骨骼肌中的一种多催化蛋白酶含有催化苯甲酰 - 缬氨酰 - 甘氨酰 - 精氨酸4 - 甲基 - 7 - 香豆素酰胺、琥珀酰 - 丙氨酰 - 丙氨酰 - 苯丙氨酸4 - 甲基香豆素酰胺和[14C]甲基酪蛋白以及苄氧羰基 - 亮氨酰 - 亮氨酰 - 谷氨酸2 - 萘基酰胺降解的活性位点。这些活性被1 mM十二烷基硫酸钠激活7 - 14倍。这种激活导致对蛋白酶抑制剂抑肽酶的敏感性更高,以及被针对未激活酶产生的抗体抑制和沉淀的能力更低。由于在SDS激活形式中未观察到分子量或亚基组成的变化,在激活步骤中似乎发生了一些构象变化。在生理浓度的脂肪酸存在下观察到更明显的激活;100 microM浓度的油酸刺激蛋白酶约50倍。与未激活的蛋白酶相比,激活的酶在体外能显著降解肌肉细胞质蛋白。因此,在体内,诸如脂肪酸等潜在激活剂能够诱导多催化蛋白酶参与肌肉蛋白分解,这并非不可能。