Amino acid residues of Escherichia coli acyl carrier protein involved in heterologous protein interactions.
Amino acid residues of Escherichia coli acyl carrier protein involved in heterologous protein interactions.
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DOI：
10.1021/bi0261950
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发表时间：
2003-01
期刊：
影响因子：
2.9
通讯作者：
L. Worsham;L. Earls;C. Jolly;Keisha G Langston;M. Trent;M. L. Ernst-Fonberg
中科院分区：
文献类型：
--
作者：
L. Worsham;L. Earls;C. Jolly;Keisha G Langston;M. Trent;M. L. Ernst-Fonberg
酰基载体蛋白（ACP）是一种高度保守的小蛋白，在植物和细菌的脂质代谢过程中与多种蛋白质相互作用，在许多反应中发挥重要作用。ACP的蛋白质部分与其许多相互作用蛋白质之间的正确识别和精确对齐的性质尚不清楚。来自许多植物和细菌的ACP之间保守的残基被认为是可能是至关重要的ACP的功能，包括蛋白质-蛋白质相互作用，并确定ACP的表面上的高疏水性的氨基酸残基簇的方法被用来估计可能参与特定的ACP-蛋白质相互作用的残基。在此信息的基础上，多个残基的单位点突变分析，一次一个，ACP被用来探测潜在的接触残基的身份ACPSH或酰基-ACP参与特定的相互作用与选定的酶。特定的ACP残基的作用更精确地定义后，与大肠杆菌溶血素激活酰基转移酶，HlyC的特异性相互作用的各种肉豆蔻酰突变型ACP的定点荧光分析。这是通过选择性标记每个突变位点，一次一个，用环境敏感的荧光探针，并观察其荧光行为的存在和不存在的HlyC。因此，ACP参与选定的大分子相互作用的一部分的图片已经出现。
Acyl carrier protein (ACP) is a small, highly conserved protein with an essential role in a myriad of reactions throughout lipid metabolism in plants and bacteria where it interacts with a remarkable diversity of proteins. The nature of the proper recognition and precise alignment between the protein moieties of ACP and its many interactive proteins is not understood. Residues conserved among ACPs from numerous plants and bacteria were considered as possibly being crucial to ACP's function, including protein-protein interaction, and a method of identifying amino acid residue clusters of high hydrophobicity on ACP's surface was used to estimate residues possibly involved in specific ACP-protein interactions. On the basis of this information, single-site mutation analysis of multiple residues, one at a time, of ACP was used to probe the identities of potential contact residues of ACPSH or acyl-ACP involved in specific interactions with selected enzymes. The roles of particular ACP residues were more precisely defined by site-directed fluorescence analyses of various myristoyl-mutant-ACPs upon specific interaction with the Escherichia coli hemolysin-activating acyltransferase, HlyC. This was done by selectively labeling each mutated site, one at a time, with an environmentally sensitive fluoroprobe and observing its fluorescence behavior in the absence and presence of HlyC. Consequently, a picture of the portion of ACP involved in selected macromolecular interaction has emerged.