Biochemical evidence for conformational changes in the cross‐talk between adenylation and peptidyl‐carrier protein domains of nonribosomal peptide synthetases
Biochemical evidence for conformational changes in the cross‐talk between adenylation and peptidyl‐carrier protein domains of nonribosomal peptide synthetases
复制标题
非核糖体肽合成酶的腺苷酸化和肽基载体蛋白结构域之间的串扰构象变化的生化证据
DOI：
10.1111/j.1742-4658.2009.07551.x
复制
发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Zettler
中科院分区：
文献类型：
--
作者：
Zettler
非核糖体多肽合成酶可作为多结构域蛋白质模板，用于生产大量具有重要药用价值的天然产物。为了正确组装所需的天然产物，必须在空间和时间水平上精确地控制不同催化中心和与肽基载体蛋白结合的反应中间体之间的相互作用。我们通过部分胰酶切、天然PAGE和凝胶过滤分析，以及化学标记实验，研究了甘草素S合成酶I(EC 5.1.1.11)中腺苷(A)结构域与肽载体蛋白的相互作用。我们的数据表明，多肽载体蛋白的4‘-磷酸丙氨酸部分由于A 结构域的构象变化而改变其位置，这是由氨基酰基腺苷模拟物结合引起的。在氨基转移到4‘-磷酸天冬氨酸部分的阶段，两个结构域之间的有效相互作用伴随着高度致密的全息蛋白构象。这些结果为多域非核糖体肽合成酶的A和肽载体蛋白结构域之间的构象变化提供了第一个生化证据。
Nonribosomal peptide synthetases serve as multidomain protein templates for producing a wealth of pharmaceutically important natural products. For the correct assembly of the desired natural product the interactions between the different catalytic centres and the reaction intermediates bound to the peptidyl carrier protein must be precisely controlled at spatial and temporal levels. We have investigated the interplay between the adenylation (A) domain and the peptidyl carrier protein in the gramicidin S synthetase I (EC 5.1.1.11) via partial tryptic digests, native PAGE and gel‐filtration analysis, as well as by chemical labeling experiments. Our data imply that the 4′‐phosphopantetheine moiety of the peptidyl carrier protein changes its position as a result of a conformational change in the A domain, which is induced by the binding of an amino acyl adenylate mimic. The productive interaction between the two domains at the stage of the amino acyl transfer onto the 4′‐phosphopantetheine moiety is accompanied by a highly compact protein conformation of the holo‐protein. These results provide the first biochemical evidence for the occurrence of conformational changes in the cross‐talk between A and peptidyl carrier protein domains of a multidomain nonribosomal peptide synthetase.
登录
查看更多内容
影响因子：
4.8
作者：
STACHELHAUS, T;MARAHIEL, MA
通讯作者：
MARAHIEL, MA
影响因子：
15
作者：
Lai, Jonathan R.;Fischbach, Michael A.;Walsh, Christopher T.
通讯作者：
Walsh, Christopher T.
影响因子：
5.7
作者：
Bruner, SD;Weber, T;Stubbs, MT
通讯作者：
Stubbs, MT
影响因子：
14.8
作者：
Ferreras, JA;Ryu, JS;Quadri, LEN
通讯作者：
Quadri, LEN
影响因子：
5.6
作者：
Samel, Stefan A.;Wagner, Bjoern;Essen, Lars-Oliver
通讯作者：
Essen, Lars-Oliver