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SMc01553 是苜蓿中华根瘤菌 1021 中的第六个酰基辅酶 A 蛋白。

SMc01553 is the sixth acyl carrier protein in Sinorhizobium meliloti 1021.

机构信息

Programa de Ecología Genómica, Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Apdo. Postal 565-A, Cuernavaca, Morelos, C.P. 62251, Mexico.

Programa de Genómica Funcional de Procariontes, Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Apdo. Postal 565-A, Cuernavaca, Morelos, C.P. 62251, Mexico.

出版信息

Microbiology (Reading). 2010 Jan;156(Pt 1):230-239. doi: 10.1099/mic.0.033480-0. Epub 2009 Oct 1.

Abstract

Acyl carrier proteins (ACPs) are required for the transfer of acyl intermediates during fatty acid and polyketide syntheses. In Sinorhizobium meliloti 1021 there are five known ACPs: AcpP, NodF, AcpXL, the ACP domain in RkpA and SMb20651. The genome sequence of S. meliloti 1021 also reveals the ORF SMc01553, annotated as a putative ACP. smc01553 is part of a 6.6 kb DNA region that is duplicated in the chromosome and in the pSymb plasmid, the result of a recent duplication event. SMc01553 overexpressed in Escherichia coli was labelled in vivo with [(3)H]beta-alanine, a biosynthetic building block of the 4'-phosphopantetheine prosthetic group of ACPs. The purified SMc01553 was modified with 4'-phosphopantetheine in the presence of S. meliloti holo-ACP synthase, and this modification resulted in a major conformational change of the protein structure, since the holo-form runs faster in native PAGE than the apo-form. SMc01553 could not be loaded with a malonyl group by malonyl-CoA-ACP transacylase from S. meliloti. Using RT-PCR we could show the presence of mRNA for SMc01553 and of the duplicated ORF SMb22007 in cultures of S. meliloti. However, a mutant in which the two duplicated regions were deleted did not show any different phenotype with respect to the wild-type in the free-living or symbiotic lifestyle.

摘要

酰基载体蛋白 (ACP) 是脂肪酸和聚酮合成过程中酰基中间体转移所必需的。在根瘤菌属中,有 5 种已知的 ACP:AcpP、NodF、AcpXL、RkpA 中的 ACP 结构域和 SMb20651。根瘤菌属 1021 的基因组序列还揭示了 ORF SMc01553,被注释为一个假定的 ACP。smc01553 是一个 6.6kb 的 DNA 区域的一部分,该区域在染色体和 pSymb 质粒中重复,这是最近发生的重复事件的结果。SMc01553 在大肠杆菌中过表达,并用 [(3)H]β-丙氨酸进行体内标记,这是 ACP 4'-磷酸泛酰巯基乙胺辅基的生物合成构建块。在含有根瘤菌属全酶 ACP 合酶的情况下,纯化的 SMc01553 被 4'-磷酸泛酰巯基乙胺修饰,这导致蛋白质结构发生重大构象变化,因为全酶形式在天然 PAGE 中比脱辅基形式跑得更快。SMc01553 不能被根瘤菌属的丙二酰基-CoA-ACP 转酰基酶加载丙二酰基。使用 RT-PCR,我们可以证明 SMc01553 的 mRNA 和复制的 ORF SMb22007 在根瘤菌属的培养物中存在。然而,在自由生活或共生生活方式中,缺失这两个重复区域的突变体与野生型相比没有表现出任何不同的表型。

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