Computational design of enone-binding proteins with catalytic activity for the Morita-Baylis-Hillman reaction.
Computational design of enone-binding proteins with catalytic activity for the Morita-Baylis-Hillman reaction.
复制标题
DOI：
10.1021/cb3006227
复制
发表时间：
2013-04-19
影响因子：
4
通讯作者：
Baker, David
中科院分区：
文献类型：
--
作者：
Bjelic, Sinisa;Nivon, Lucas G.;Celebi-Oelcuem, Nihan;Kiss, Gert;Rosewall, Carolyn F.;Lovick, Helena M.;Ingalls, Erica L.;Gallaher, Jasmine Lynn;Seetharaman, Jayaraman;Lew, Scott;Montelione, Gaetano Thomas;Hunt, John Francis;Michael, Forrest Edwin;Houk, K. N.;Baker, David
The Morita-Baylis-Hillman reaction forms a carbon-carbon bond between the alpha carbon of a conjugated carbonyl compound and a carbon electrophile. The reaction mechanism involves Michael addition of a nucleophile catalyst at the carbonyl beta carbon, followed by bond formation with the electrophile and catalyst disassociation to release the product. We used Rosetta to design 48 proteins containing active sites predicted to carry out this mechanism, of which two show catalytic activity by mass spectrometry (MS). Substrate labeling measured by MS and site-directed mutagenesis experiments show that the designed active-site residues are responsible for activity, although rate acceleration over background is modest. To characterize the designed proteins, we developed a fluorescence-based screen for intermediate formation in cell lysates, carried out microsecond molecular dynamics simulations, and solved X-ray crystal structures. These data indicate a partially formed active site, and suggest several clear avenues for designing more active catalysts.
登录
查看更多内容
影响因子：
3.7
作者：
Richter F;Leaver-Fay A;Khare SD;Bjelic S;Baker D
通讯作者：
Baker D
影响因子：
1.8
作者：
Iwabuchi, Y;Sugihara, T;Hatakeyama, S
通讯作者：
Hatakeyama, S
影响因子：
8
作者：
Arai, R;Kukimoto-Niino, M;Yokoyama, S
通讯作者：
Yokoyama, S
影响因子：
1.8
作者：
Reetz, Manfred T.;Mondiere, Regis;Carballeira, Jose Daniel
通讯作者：
Carballeira, Jose Daniel
影响因子：
56.9
作者：
Savile, Christopher K.;Janey, Jacob M.;Hughes, Gregory J.
通讯作者：
Hughes, Gregory J.