Rauwerdink Alissa, Kazlauskas Romas J
University of Minnesota, Department of Biochemistry, Molecular Biology & Biophysics and The Biotechnology Institute, Saint Paul, MN 55108 USA.
ACS Catal. 2015 Oct 2;5(10):6153-6176. doi: 10.1021/acscatal.5b01539. Epub 2015 Sep 9.
Enzymes within a family often catalyze different reactions. In some cases, this variety stems from different catalytic machinery, but in other cases the machinery is identical; nevertheless, the enzymes catalyze different reactions. In this review, we examine the subset of α/β-hydrolase fold enzymes that contain the serine-histidine-aspartate catalytic triad. In spite of having the same protein fold and the same core catalytic machinery, these enzymes catalyze seventeen different reaction mechanisms. The most common reactions are hydrolysis of C-O, C-N and C-C bonds (Enzyme Classification (EC) group 3), but other enzymes are oxidoreductases (EC group 1), acyl transferases (EC group 2), lyases (EC group 4) or isomerases (EC group 5). Hydrolysis reactions often follow the canonical esterase mechanism, but eight variations occur where either the formation or cleavage of the acyl enzyme intermediate differs. The remaining eight mechanisms are lyase-type elimination reactions, which do not have an acyl enzyme intermediate and, in four cases, do not even require the catalytic serine. This diversity of mechanisms from the same catalytic triad stems from the ability of the enzymes to bind different substrates, from the requirements for different chemical steps imposed by these new substrates and, only in about half of the cases, from additional hydrogen bond partners or additional general acids/bases in the active site. This detailed analysis shows that binding differences and non-catalytic residues create new mechanisms and are essential for understanding and designing efficient enzymes.
同一家族中的酶通常催化不同的反应。在某些情况下,这种多样性源于不同的催化机制,但在其他情况下,机制是相同的;然而,这些酶催化不同的反应。在本综述中,我们研究了包含丝氨酸 - 组氨酸 - 天冬氨酸催化三联体的α/β-水解酶折叠酶子集。尽管具有相同的蛋白质折叠和相同的核心催化机制,但这些酶催化十七种不同的反应机制。最常见的反应是C - O、C - N和C - C键的水解(酶分类(EC)第3组),但其他酶是氧化还原酶(EC第1组)、酰基转移酶(EC第2组)、裂合酶(EC第4组)或异构酶(EC第5组)。水解反应通常遵循典型的酯酶机制,但存在八种变体,其中酰基酶中间体的形成或裂解有所不同。其余八种机制是裂合酶型消除反应,它们没有酰基酶中间体,并且在四种情况下甚至不需要催化丝氨酸。来自相同催化三联体的这种机制多样性源于酶结合不同底物的能力、这些新底物对不同化学步骤的要求,并且仅在大约一半的情况下,源于活性位点中额外的氢键伙伴或额外的广义酸/碱。这种详细分析表明,结合差异和非催化残基创造了新的机制,对于理解和设计高效酶至关重要。