配体(生物化学)
化学
生物物理学
互补性(分子生物学)
蛋白质设计
结合位点
血浆蛋白结合
蛋白质工程
立体化学
蛋白质结构
结晶学
计算生物学
生物化学
生物
遗传学
受体
酶
作者
Austin L. Day,Per Greisen,Lindsey Doyle,Alberto Schena,Nephi Stella,Kai Johnsson,David Baker,Barry Stoddard
出处
期刊:Protein Engineering Design & Selection
[Oxford University Press]
日期:2018-10-01
卷期号:31 (10): 375-387
被引量:6
标识
DOI:10.1093/protein/gzy031
摘要
Abstract Attempts to create novel ligand-binding proteins often focus on formation of a binding pocket with shape complementarity against the desired ligand (particularly for compounds that lack distinct polar moieties). Although designed proteins often exhibit binding of the desired ligand, in some cases they display unintended recognition behavior. One such designed protein, that was originally intended to bind tetrahydrocannabinol (THC), was found instead to display binding of 25-hydroxy-cholecalciferol (25-D3) and was subjected to biochemical characterization, further selections for enhanced 25-D3 binding affinity and crystallographic analyses. The deviation in specificity is due in part to unexpected altertion of its conformation, corresponding to a significant change of the orientation of an α-helix and an equally large movement of a loop, both of which flank the designed ligand-binding pocket. Those changes led to engineered protein constructs that exhibit significantly more contacts and complementarity towards the 25-D3 ligand than the initial designed protein had been predicted to form towards its intended THC ligand. Molecular dynamics simulations imply that the initial computationally designed mutations may contribute to the movement of the helix. These analyses collectively indicate that accurate prediction and control of backbone dynamics conformation, through a combination of improved conformational sampling and/or de novo structure design, represents a key area of further development for the design and optimization of engineered ligand-binding proteins.
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