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光合能量守恒中的巯基。V. 邻碘代苯甲酸在菠菜叶绿体偶联因子中形成的新二硫键的定位

Sulphydryl groups in photosynthetic energy conservation. V. Localization of the new disulfide bridges formed by o-iodosobenzoate in coupling factor of spinach chloroplasts.

作者信息

Andreo C S, Ravizzini R A, Vallejos R H

出版信息

Biochim Biophys Acta. 1979 Aug 14;547(2):370-9. doi: 10.1016/0005-2728(79)90018-5.

Abstract
  1. Chemical modification by o-iodosobenzoate of soluble chloroplast coupling factor 1 (CF1) during heat activation resulted in inhibition of its Ca-ATPase activity and in the formation of two new intrapeptide disulfide bridges as suggested by: (a) the disappearance of three out of four accessible thiol groups, two from gamma and one from a beta subunit as a consequence of CF1 modification by o-iodosobenzoate; (b) the total free sulphydryl groups of CF1 were reduced from 8 to 4 after modification of CF1 by o-iodosobenzoate. Two groups disappeared from beta and two from gamma subunits; (c) a second heating step of CF1 in the presence of 10 mM dithioerythritol reversed the inhibition of the ATPase and reduced both the newly formed disulfide bridges and those present in native CF1. 2. Modification of chloroplasts in the light with o-iodosobenzoate resulted in the inhibition of photophosphorylation and ATPase. CF1 isolated and purified from these chloroplasts had its Ca-ATPase activity inhibited and two new disulfide bridges. The total number of free sulphydryl groups was reduced from 8 to 4 and three accessible groups disappeared from beta and gamma subunits.
摘要
  1. 在热激活过程中,邻碘代苯甲酸对可溶性叶绿体偶联因子1(CF1)进行化学修饰,导致其钙ATP酶活性受到抑制,并形成了两个新的肽内二硫键,这由以下几点表明:(a)四个可及巯基中的三个消失,其中两个来自γ亚基,一个来自β亚基,这是邻碘代苯甲酸对CF1进行修饰的结果;(b)CF1经邻碘代苯甲酸修饰后,其总游离巯基从8个减少到4个。两个基团从β亚基消失,两个从γ亚基消失;(c)在10 mM二硫赤藓糖醇存在下,CF1的第二步加热逆转了ATP酶的抑制作用,并减少了新形成的二硫键以及天然CF1中存在的二硫键。2. 用邻碘代苯甲酸在光照下对叶绿体进行修饰,导致光合磷酸化和ATP酶受到抑制。从这些叶绿体中分离纯化得到的CF1,其钙ATP酶活性受到抑制,并且有两个新的二硫键。游离巯基总数从8个减少到4个,三个可及基团从β亚基和γ亚基消失。

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