The mechanism of α1-antitrypsin polymerization probed by fluorescence spectroscopy
The mechanism of α1-antitrypsin polymerization probed by fluorescence spectroscopy
复制标题
DOI：
10.1006/abbi.1998.0751
复制
发表时间：
1998-08-15
影响因子：
3.9
通讯作者：
Bottomley, SP
中科院分区：
文献类型：
--
作者：
James, EL;Bottomley, SP
肝细胞内α(1)-抗胰蛋白酶的聚合导致α(1)-抗胰蛋白酶缺乏。人们对聚合过程的构象变化和动力学知之甚少。在这里，我们描述了使用荧光探针 4,4'-二苯胺基-1,1'-联萘-5,5'-二磺酸盐 (bis-ANS) 研究聚合反应的荧光实验，该探针与天然和聚合的 α(1)-抗胰蛋白酶结合。在 α(1)-抗胰蛋白酶聚合物的形成过程中观察到双-ANS 荧光的双相变化。最初观察到荧光信号快速增加；随后荧光信号逐渐减弱。第一阶段是构象变化，其中 α(1)-抗胰蛋白酶的 A β-折叠打开，而第二阶段表示反应中心环插入另一个分子的 A β-折叠，因此决定聚合过程的速率。 (C) 1998 年学术出版社。
The polymerization of alpha(1)-antitrypsin within the hepatic cell leads to alpha(1)-antitrypsin deficiency. Both the conformational changes and the kinetics of the polymerization process are poorly understood. Here we describe fluorescence experiments investigating the polymerization reaction using the fluorescent probe 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonate (bis-ANS) which bound to both native and polymerized alpha(1)-antitrypsin. Biphasic changes in bis-ANS fluorescence were observed during formation of alpha(1)-antitrypsin polymers. Initially a rapid increase in fluorescence signal was observed; it was followed by a gradual reduction in fluorescence signal. The first phase is a conformational change in which the A beta-sheet of alpha(1)-antitrypsin opens, whereas the second phase represents the insertion of the reactive center loop into the A beta-sheet of another molecule and therefore determines the rate of the polymerization process. (C) 1998 Academic Press.