Computationally mapping sequence space to understand evolutionary protein engineering

被引:5
作者
Armstrong, Kathryn A. [1 ]
Tidor, Bruce [1 ,2 ]
机构
[1] MIT, Dept Biol Engn, Comp Sci & Artificial Intelligence Lab, Cambridge, MA 02139 USA
[2] MIT, Dept Elect Engn & Comp Sci, Cambridge, MA 02139 USA
关键词
D O I
10.1021/bp070134h
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Evolutionary protein engineering has been dramatically successful, producing a wide variety of new proteins with altered stability, binding affinity, and enzymatic activity. However, the success of such procedures is often unreliable, and the impact of the choice of protein, engineering goal, and evolutionary procedure is not well understood. We have created a framework for understanding aspects of the protein engineering process by computationally mapping regions of feasible sequence space for three small proteins using structure-based design protocols. We then tested the ability of different evolutionary search strategies to explore these sequence spaces. The results point to a non-intuitive relationship between the error-prone PCR mutation rate and the number of rounds of replication. The evolutionary relationships among feasible sequences reveal hub-like sequences that serve as particularly fruitful starting sequences for evolutionary search. Moreover, genetic recombination procedures were examined, and tradeoffs relating sequence diversity and search efficiency were identified: This framework allows us to consider the impact of protein structure on the allowed sequence space and therefore on the challenges that each protein presents to error-prone PCR and genetic recombination procedures.
引用
收藏
页码:62 / 73
页数:12
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