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胰蛋白酶处理大鼠肾刷状缘膜对氨基酸转运的影响。

The effect on amino acid transport of trypsin treatment of rat renal brush border membranes.

作者信息

Hsu B Y, Corcoran S M, Marshall C M, Segal S

出版信息

Biochim Biophys Acta. 1982 Jul 28;689(2):181-93. doi: 10.1016/0005-2736(82)90250-4.

Abstract

Trypsin treatment of isolated rat renal brush border membrane vesicles which preferentially releases L-leucine aminopeptidase (EC 3.4.11.2) decreases their ability to take up a variety of amino acids under Na+ -gradient conditions. Such treatment did not alter the osmotic properties of the vesicles nor affect their fragility. A linear correlation could be demonstrated between the L-leucine aminopeptidase activity of the membranes and the initial rate of uptake of L-leucine and L-proline. Velocity of uptake-concentration dependence studies with these substrates indicate that the major effect of trypsinization is to decrease the maximum velocity (Vmax1) of the low-Km high-affinity system with little effect on the Vmax2 of the high-Km low-affinity transport process and no effect on the apparent Michaelis constants of either. Although the data indicate that L-leucine aminopeptidase activity and uptake of l-leucine and L-proline are affected in parallel, they should not be construed to imply a role of the enzyme in the transport process, especially in view of the global decrease in the uptake of various amino acids and sugars.

摘要

用胰蛋白酶处理分离出的大鼠肾刷状缘膜囊泡(这种处理优先释放L-亮氨酸氨肽酶,即EC 3.4.11.2),会降低它们在Na⁺梯度条件下摄取多种氨基酸的能力。这种处理不会改变囊泡的渗透特性,也不会影响其脆性。膜的L-亮氨酸氨肽酶活性与L-亮氨酸和L-脯氨酸的初始摄取速率之间可以证明存在线性相关性。对这些底物进行摄取速度-浓度依赖性研究表明,胰蛋白酶处理的主要作用是降低低Km高亲和力系统的最大速度(Vmax1),对高Km低亲和力转运过程的Vmax2影响很小,对两者的表观米氏常数均无影响。尽管数据表明L-亮氨酸氨肽酶活性以及L-亮氨酸和L-脯氨酸的摄取受到平行影响,但不应将其解释为暗示该酶在转运过程中起作用,特别是考虑到各种氨基酸和糖的摄取普遍下降。

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