Crystal structure of the surfactin synthetase-activating enzyme Sfp:: a prototype of the 4′-phosphopantetheinyl transferase superfamily
Crystal structure of the surfactin synthetase-activating enzyme Sfp:: a prototype of the 4′-phosphopantetheinyl transferase superfamily
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DOI：
10.1093/emboj/18.23.6823
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发表时间：
1999-12-01
期刊：
影响因子：
11.4
通讯作者：
Ficner, R
中科院分区：
文献类型：
--
作者：
Reuter, K;Mofid, MR;Ficner, R
枯草芽孢杆菌Sfp蛋白通过将辅酶A（CoA）的4 '-磷酸泛酰巯基乙胺基部分转移到所有PCP中保守的丝氨酸残基来激活表面活性素合成酶的肽基载体蛋白（PCP）结构域。其广泛的PCP底物谱使Sfp成为用于组合非核糖体肽合成的生物技术上有价值的酶。在1.8埃分辨率下测定的SfpCoA复合物的结构揭示了一种新的α/β折叠，其表现出意想不到的分子内2重假对称性。这表明同二聚磷酸泛酰巯基乙胺基转移酶如酰基载体蛋白合酶的类似折叠和二聚化模式。Sfp的活性位点容纳镁离子，其与CoA焦磷酸盐复合，三个酸性氨基酸和一个水分子的侧链。CoA的结合方式在许多方面不同于所有已知的CoA-蛋白质复合物结构。该结构揭示了可能参与与五氯苯酚底物相互作用的区域。
The Bacillus subtilis Sfp protein activates the peptidyl carrier protein (PCP) domains of surfactin synthetase by transferring the 4'-phosphopantetheinyl moiety of coenzyme A (CoA) to a serine residue conserved in all PCPs. Its wide PCP substrate spectrum renders Sfp a biotechnologically valuable enzyme for use in combinatorial non-ribosomal peptide synthesis. The structure of the SfpCoA complex determined at 1.8 Angstrom resolution reveals a novel alpha/beta-fold exhibiting an unexpected intramolecular 2-fold pseudosymmetry, This suggests a similar fold and dimerization mode for the homodimeric phosphopantetheinyl transferases such as acyl carrier protein synthase, The active site of Sfp accommodates a magnesium ion, which is complexed by the CoA pyrophosphate, the side chains of three acidic amino acids and one water molecule. CoA is bound in a fashion that differs in many aspects from all known CoA-protein complex structures. The structure reveals regions likely to be involved in the interaction with the PCP substrate.