The role of distant mutations and allosteric regulation on LovD active site dynamics.
The role of distant mutations and allosteric regulation on LovD active site dynamics.
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DOI：
10.1038/nchembio.1503
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发表时间：
2014-06
影响因子：
14.8
通讯作者：
Houk, K. N.
中科院分区：
文献类型：
--
作者：
Jimenez-Oses, Gonzalo;Osuna, Silvia;Gao, Xue;Sawaya, Michael R.;Gilson, Lynne;Collier, Steven J.;Huisman, Gjalt W.;Yeates, Todd O.;Tang, Yi;Houk, K. N.
Natural enzymes have evolved to perform their cellular functions under complex selective pressures, which often require their catalytic activities to be regulated by other proteins. We contrasted a natural enzyme, LovD, which acts on a protein-bound (LovF) acyl substrate, with a laboratory-generated variant that was transformed by directed evolution to accept instead a small free acyl thioester, and no longer requires the acyl carrier protein. The resulting 29-mutant variant is 1000-fold more efficient in the synthesis of the drug simvastatin than the wild-type LovD. This is the first non-patent report of the enzyme currently used for the manufacture of simvastatin, as well as the intermediate evolved variants. Crystal structures and microsecond molecular dynamics simulations revealed the mechanism by which the laboratory-generated mutations free LovD from dependence on protein-protein interactions. Mutations dramatically altered conformational dynamics of the catalytic residues, obviating the need for allosteric modulation by the acyl carrier LovF.
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DOI：
10.1126/science.1198542
发表时间：
2011-04-08
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Bhabha G;Lee J;Ekiert DC;Gam J;Wilson IA;Dyson HJ;Benkovic SJ;Wright PE
通讯作者：
Wright PE
DOI：
10.1073/pnas.0408343102
发表时间：
2005-05-10
影响因子：
11.1
作者：
Wong, KF;Selzer, T;Hammes-Schiffer, S
通讯作者：
Hammes-Schiffer, S
影响因子：
46.9
作者：
Fox, Richard J.;Davis, S. Christopher;Huisman, Gjalt W.
通讯作者：
Huisman, Gjalt W.
DOI：
10.1073/pnas.1207028109
发表时间：
2012-10-23
影响因子：
11.1
作者：
Agarwal, Vinayak;Lin, Steven;Cronan, John E.
通讯作者：
Cronan, John E.
影响因子：
56.9
作者：
Shaw, David E.;Maragakis, Paul;Wriggers, Willy
通讯作者：
Wriggers, Willy