期刊
FEBS JOURNAL
卷 282, 期 23, 页码 4538-4547出版社
WILEY
DOI: 10.1111/febs.13513
关键词
crystal structure; gating mechanism; inhibition; lid; lipase
资金
- National High Technology Research and Development Program of China [2014AA093514]
- National Science Funds for Excellent Youth Scholars [31222043]
- National Natural Science Foundation of China [21406076]
- Fundamental Research Funds for the Central Universities [2015ZM059]
- Science and Technology Planning Project of Guangdong Province [2014B020204003]
- Natural Science Foundation of Guangdong Province of China [2015A030310381]
- National Science Foundation for Young Scholars of China [31500638]
Monoacylglycerol and diacylglycerol lipases are industrially interesting enzymes, due to the health benefits that arise from the consumption of diglycerides compared to the traditional triglyceride oils. Most lipases possess an alpha-helix (lid) directly over the catalytic pocket which regulates the activity of the enzyme. Generally, lipases exist in active and inactive conformations, depending on the positioning of this lid subdomain. However, lipase SMG1, a monoacylglycerol and diacylglycerol specific lipase, has an atypical activation mechanism. In the present study we were able to prove by crystallography, in silico analysis and activity tests that only two positions, residues 102 and 278, are responsible for a gating mechanism that regulates the active and inactive states of the lipase, and that no significant structural changes take place during activation except for oxyanion hole formation. The elucidation of the gating effect provided data enabling the rational design of improved lipases with 6-fold increase in the hydrolytic activity toward diacylglycerols, just by providing additional substrate stabilization with a single mutation (F278N or F278T). Due to the conservation of F278 among the monoacylglycerol and diacylglycerol lipases in the Rhizomucor miehei lipase-like family, the gating mechanism described herein might represent a general mechanism applicable to other monoacylglycerol and diacylglycerol lipases as well.
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