嘌呤核苷酸生物合成途径中加氮酶的趋同进化——基于腺苷琥珀酸合成酶(PurA)的结构分析。

IF 0.8 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of General and Applied Microbiology Pub Date : 2023-11-15 Epub Date: 2023-06-10 DOI:10.2323/jgam.2023.05.002
Gen-Ichi Sampei, Hironori Ishii, Hiroyuki Taka, Gota Kawai
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引用次数: 0

摘要

腺苷琥珀酸合成酶(PurA)是一种在嘌呤核苷酸生物合成途径中由天冬氨酸向单磷酸肌苷(IMP)添加氮的酶。然后延胡索酸被琥珀酸腺苷裂解酶(PurB)除去,留下一个氨基。另外还有两种酶可以催化类似PurA的天冬氨酸加成反应,一种是嘌呤核苷酸生物合成途径(SAICAR合成酶,PurC),另一种是精氨酸生物合成途径(精氨酸琥珀酸合成酶,ArgG)。为了研究这些加氮酶的来源,我们对来自Thermus thermophilus HB8的PurA (TtPurA)进行了纯化和结晶,并以2.10 Å的分辨率测定了与IMP络合的晶体结构。TtPurA具有同二聚体结构,在二聚体界面,一个亚基的Arg135与与另一个亚基结合的IMP相互作用,表明IMP结合有助于二聚体的稳定性。在TtPurA和EcPurA中,His41侧链的不同构象表明,His41侧链翻转可能对GTP的γ-磷酸在IMP第6位靠近氧的位置定向以接受亲核攻击起重要作用。此外,通过PurA、PurC和ArgG的三维结构和活性位点的比较,可以看出PurA和PurC的活性位点趋近于相似的结构,进行相似的反应。
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Convergent evolution of nitrogen-adding enzymes in the purine nucleotide biosynthetic pathway, based on structural analysis of adenylosuccinate synthetase (PurA).

Adenylosuccinate synthetase (PurA) is an enzyme responsible for the nitrogen addition to inosine monophosphate (IMP) by aspartate in the purine nucleotide biosynthetic pathway. And after which the fumarate is removed by adenylosuccinate lyase (PurB), leaving an amino group. There are two other enzymes that catalyze aspartate addition reactions similar to PurA, one in the purine nucleotide biosynthetic pathway (SAICAR synthetase, PurC) and the other in the arginine biosynthetic pathway (argininosuccinate sythetase, ArgG). To investigate the origin of these nitrogen-adding enzymes, PurA from Thermus thermophilus HB8 (TtPurA) was purified and crystallized, and crystal structure complexed with IMP was determined with a resolution of 2.10 Å. TtPurA has a homodimeric structure, and at the dimer interface, Arg135 of one subunit interacts with the IMP bound to the other subunit, suggesting that IMP binding contributes to dimer stability. The different conformation of His41 side chain in TtPurA and EcPurA suggests that side chain flipping of the His41 might play an important role in orienting γ-phosphate of GTP close to oxygen at position 6 of IMP, to receive the nucleophilic attack. Moreover, through comparison of the three-dimensional structures and active sites of PurA, PurC, and ArgG, it was suggested that the active sites of PurA and PurC converged to similar structures for performing similar reactions.

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来源期刊
Journal of General and Applied Microbiology
Journal of General and Applied Microbiology 生物-生物工程与应用微生物
CiteScore
2.40
自引率
0.00%
发文量
42
审稿时长
6-12 weeks
期刊介绍: JGAM is going to publish scientific reports containing novel and significant microbiological findings, which are mainly devoted to the following categories: Antibiotics and Secondary Metabolites; Biotechnology and Metabolic Engineering; Developmental Microbiology; Environmental Microbiology and Bioremediation; Enzymology; Eukaryotic Microbiology; Evolution and Phylogenetics; Genome Integrity and Plasticity; Microalgae and Photosynthesis; Microbiology for Food; Molecular Genetics; Physiology and Cell Surface; Synthetic and Systems Microbiology.
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