Catalytic promiscuity in a protein superfamily

蛋白质超家族中的催化混杂

• 批准号: BB/I004327/1
• 负责人: Florian Hollfelder
• 金额: $ 58.55万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2011
• 资助国家: 英国
• 起止时间: 2011 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FI004327%2F1
• 关键词: Catalytic; promiscuity; protein; superfamily

Understanding of enzyme catalysis remains a daunting challenge, despite intense research efforts in basic and applied research. Our understanding certainly fails the most severe test - that of making catalysts that meet the efficiency of natural enzymes. This proposal addresses the determinants of catalytic efficiency in hydrolytic enzymes, using as a test bed a superfamily of catalysts that are connected by structural homology as members of the alkaline phosphatase superfamily as well as often reciprocal and crosswise catalytic promiscuity. Starting from our recent identification of one remarkable example for multiple promiscuity and high efficiency (Proc. Natl. Acad. Sci - in press, doi/10.1073/pnas.0903951107), we propose to address the factors that determine selectivity and efficiency, by structural and kinetic analysis and by directed evolution.

尽管在基础和应用研究方面进行了大量的研究工作，但对酶催化的理解仍然是一个艰巨的挑战。我们的理解当然没有通过最严格的测试--即制造符合天然酶效率的催化剂。这个建议解决了水解酶的催化效率的决定因素，作为一个测试床的催化剂的超家族，连接的碱性磷酸酶超家族的成员，以及经常相互和相互催化混杂的结构同源性。从我们最近发现的一个关于多重滥交和高效率的显着例子开始（Proc. Natl. Acad. Sci - in Press，doi/10.1073/pnas.0903951107），我们提出通过结构和动力学分析以及通过定向进化来解决决定选择性和效率的因素。

项目成果

Specificity Effects of Amino Acid Substitutions in Promiscuous Hydrolases: Context-Dependence of Catalytic Residue Contributions to Local Fitness Landscapes in Nearby Sequence Space.

• DOI: 10.1002/cbic.201600657
• 发表时间: 2017-06-01
• 期刊: Chembiochem : a European journal of chemical biology
• 作者: Bayer CD;van Loo B;Hollfelder F
• 通讯作者: Hollfelder F

Ultrahigh-throughput discovery of promiscuous enzymes by picodroplet functional metagenomics.

• DOI: 10.1038/ncomms10008
• 发表时间: 2015-12-07
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Colin PY;Kintses B;Gielen F;Miton CM;Fischer G;Mohamed MF;Hyvönen M;Morgavi DP;Janssen DB;Hollfelder F
• 通讯作者: Hollfelder F

Reverse evolution leads to genotypic incompatibility despite functional and active site convergence.

尽管功能和活性位点趋同，逆向进化仍会导致基因型不相容。

• DOI: 10.17863/cam.63718
• 发表时间: 2015
• 作者: Kaltenbach M

Evolutionary repurposing of a sulfatase: A new Michaelis complex leads to efficient transition state charge offset.

硫酸酯酶的进化再利用：一种新的米氏复合物导致有效的过渡态电荷抵消。

• DOI: 10.1073/pnas.1607817115
• 发表时间: 2018
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Miton CM

Specificity Effects of Amino Acid Substitutions in Promiscuous Hydrolases: Context-Dependence of Catalytic Residue Contributions to Local Fitness Landscapes in Nearby Sequence Space

混杂水解酶中氨基酸取代的特异性效应：催化残基对附近序列空间局部适应度景观贡献的上下文依赖性

• DOI: 10.17863/cam.9842
• 发表时间: 2017
• 作者: Bayer C