Revealing the hidden functional diversity of an enzyme family
Revealing the hidden functional diversity of an enzyme family
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DOI：
10.1038/nchembio.1387
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发表时间：
2014-01-01
影响因子：
14.8
通讯作者：
Salanoubat, Marcel
中科院分区：
文献类型：
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作者：
Bastard, Karine;Smith, Adam Alexander Thil;Salanoubat, Marcel
数以百万计的蛋白质数据库条目没有被分配可靠的功能，阻碍了对生物体中化学多样性的充分理解。在这里，我们描述了一个综合的策略，发现各种酶的活性催化蛋白质家族的未知或鲜为人知的功能。该方法依赖于家族内保守的通用反应的定义、代表性的高通量酶筛选、结构和建模研究以及基因组和代谢背景的分析。作为原理的证明，我们研究了DUF 849 Pfam家族，发现了14种潜在的新酶活性，从而将这些蛋白质指定为β-酮酸裂解酶。我们提出了四种酶活性在体内的作用，并建议指导进一步的功能注释的关键残基。我们的研究结果表明，家庭内的功能多样性可能在很大程度上被低估。将这种策略扩展到其他家族将提高我们对酶景观的认识。
Millions of protein database entries are not assigned reliable functions, preventing the full understanding of chemical diversity in living organisms. Here, we describe an integrated strategy for the discovery of various enzymatic activities catalyzed within protein families of unknown or little known function. This approach relies on the definition of a generic reaction conserved within the family, high-throughput enzymatic screening on representatives, structural and modeling investigations and analysis of genomic and metabolic context. As a proof of principle, we investigated the DUF849 Pfam family and unearthed 14 potential new enzymatic activities, leading to the designation of these proteins as beta-keto acid cleavage enzymes. We propose an in vivo role for four enzymatic activities and suggest key residues for guiding further functional annotation. Our results show that the functional diversity within a family may be largely underestimated. The extension of this strategy to other families will improve our knowledge of the enzymatic landscape.