Mapping of the Binding Landscape for a Picomolar Protein-Protein Complex through Computation and Experiment

被引:17
作者
Aizner, Yonatan [1 ]
Sharabi, Oz [1 ]
Shirian, Jason [1 ]
Dakwar, George R. [1 ]
Risman, Marina [1 ]
Avraham, Orly [1 ]
Shifman, Julia [1 ]
机构
[1] Hebrew Univ Jerusalem, Alexander Silberman Inst Life Sci, Dept Biol Chem, IL-91904 Jerusalem, Israel
关键词
SACCHAROMYCES-CEREVISIAE; SCANNING MUTAGENESIS; INTERACTION NETWORK; CRYSTAL-STRUCTURE; SNAKE-VENOM; ACETYLCHOLINESTERASE; DESIGN; FASCICULIN; AFFINITY; INTERFACE;
D O I
10.1016/j.str.2014.01.012
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Our understanding of protein evolution would greatly benefit from mapping of binding landscapes, i.e., changes in protein-protein binding affinity due to all single mutations. However, experimental generation of such landscapes is a tedious task due to a large number of possible mutations. Here, we use a simple computational protocol to map the binding landscape for two homologous high-affinity complexes, involving a snake toxin fasciculin and acetylcholinesterase from two different species. To verify our computational predictions, we experimentally measure binding between 25 Fas mutants and the 2 enzymes. Both computational and experimental results demonstrate that the Fas sequence is close to the optimum when interacting with its targets, yet a few mutations could further improve K-d, k(on), and k(off). Our computational predictions agree well with experimental results and generate distributions similar to those observed in other high-affinity PPIs, demonstrating the potential of simple computational protocols in capturing realistic binding landscapes.
引用
收藏
页码:636 / 645
页数:10
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