Determination of the structure of the catabolic N-succinylornithine transaminase (AstC) from Escherichia coli.

Determination of the structure of the catabolic N-succinylornithine transaminase (AstC) from Escherichia coli.
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DOI:
10.1371/journal.pone.0058298
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发表时间:
2013
期刊:
影响因子:
3.7
通讯作者:
Peat TS
Peat TS
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Newman J;Seabrook S;Surjadi R;Williams CC;Lucent D;Wilding M;Scott C;Peat TS

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大肠杆菌具有两种酰基鸟氨酸转氨酶,一种是分解代谢酶(AsTC),另一种是合成代谢酶(ArgD),参与L精氨酸代谢。虽然只有58%相同,但已证明这些酶在功能上是可互换的。在这里,我们纯化了ASTC,并获得了apo和holo-astc的X射线晶体结构以及该酶与其生理底物琥珀酰鸟氨酸的络合结构。我们将本研究中获得的结构与从其他地方获得的鼠伤寒沙门氏菌ArgD的结构进行了比较,发现了几个显著的差异。通过对接研究探讨了几种底物(鸟氨酸、琥珀酸鸟氨酸和乙酰鸟氨酸)与谷氨酸/α-酮杂化共底物的对接模式。对接研究支持了我们的观察结果,即ASTC对酰化的鸟氨酸物种比鸟氨酸本身具有强烈的偏好,并表明与酰化相关的特异性的增加是由空间位阻和脱溶效应引起的,而不是底物和酶之间的特定相互作用。
Escherichia coli possesses two acyl ornithine aminotransferases, one catabolic (AstC) and the other anabolic (ArgD), that participate in L-arginine metabolism. Although only 58% identical, the enzymes have been shown to be functionally interchangeable. Here we have purified AstC and have obtained X-ray crystal structures of apo and holo-AstC and of the enzyme complexed with its physiological substrate, succinylornithine. We compare the structures obtained in this study with those of ArgD from Salmonella typhimurium obtained elsewhere, finding several notable differences. Docking studies were used to explore the docking modes of several substrates (ornithine, succinylornithine and acetylornithine) and the co-substrate glutamate/α-ketogluterate. The docking studies support our observations that AstC has a strong preference for acylated ornithine species over ornithine itself, and suggest that the increase in specificity associated with acylation is caused by steric and desolvation effects rather than specific interactions between the substrate and enzyme.
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