Molecular Basis for Cγ-N Bond Formation by PLP-Dependent Enzyme LolC.

IF 2.9 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Biochemistry Biochemistry Pub Date : 2024-12-17 Epub Date: 2024-12-06 DOI:10.1021/acs.biochem.4c00588
Yueqi Xu, Shaonan Liu, Jinmin Gao, Yang Hai
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引用次数: 0

Abstract

Pyridoxal 5'-phosphate (PLP)-dependent enzymes catalyze a diverse array of biochemical transformations, making them invaluable biocatalytic tools for the synthesis of complex bioactive compounds. Here, we report the biochemical characterization of LolC, a PLP-dependent γ-synthase involved in the biosynthesis of loline alkaloids. LolC catalyzes the formation of a Cγ-N bond between O-acetyl--homoserine (OAH) and l-proline, generating a diamino diacid intermediate. Our findings reveal that LolC exhibits strict specificity for proline and its analogues, contrasting with the substrate promiscuity of closely related Cγ-C bond-forming enzyme Fub7. Structural analysis, using an AlphaFold model, identifies key differences in the substrate entrance tunnel of LolC, which is amphiphilic and distinct from the hydrophobic tunnel in Fub7. A mutagenesis study further highlights the functional divergence of a key active site residue between these enzymes. These results provide new insights into the substrate specificity and catalytic function of LolC, offering a valuable comparison to Fub7 and advancing our understanding of PLP-dependent enzymes involved in γ-substitution reactions.

plp依赖性酶LolC形成c - γ- n键的分子基础
吡哆醛5'-磷酸(PLP)依赖酶催化多种生化转化,使其成为合成复杂生物活性化合物的宝贵生物催化工具。在这里,我们报道了LolC的生化特性,LolC是一种plp依赖性的γ-合成酶,参与了碱生物碱的生物合成。LolC催化o -乙酰-高丝氨酸(OAH)和l-脯氨酸之间形成c - γ- n键,生成二氨基二酸中间体。我们的研究结果表明,与密切相关的c - γ- c成键酶Fub7的底物混杂性形成对比,LolC对脯氨酸及其类似物具有严格的特异性。使用AlphaFold模型进行结构分析,确定了LolC的底物入口隧道的关键差异,该隧道是两亲性的,与Fub7中的疏水隧道不同。一项诱变研究进一步强调了这些酶之间关键活性位点残基的功能差异。这些结果为LolC的底物特异性和催化功能提供了新的见解,提供了与Fub7的有价值的比较,并促进了我们对参与γ-取代反应的plp依赖性酶的理解。
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来源期刊
Biochemistry Biochemistry
Biochemistry Biochemistry 生物-生化与分子生物学
CiteScore
5.50
自引率
3.40%
发文量
336
审稿时长
1-2 weeks
期刊介绍: Biochemistry provides an international forum for publishing exceptional, rigorous, high-impact research across all of biological chemistry. This broad scope includes studies on the chemical, physical, mechanistic, and/or structural basis of biological or cell function, and encompasses the fields of chemical biology, synthetic biology, disease biology, cell biology, nucleic acid biology, neuroscience, structural biology, and biophysics. In addition to traditional Research Articles, Biochemistry also publishes Communications, Viewpoints, and Perspectives, as well as From the Bench articles that report new methods of particular interest to the biological chemistry community.
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