.png){kind=icon}
返回
Sequence coevolution and structure stabilization modulate olfactory receptor expression
.png){kind=icon}
OA
AI
DOI：10.1016/j.bpj.2022.01.015.png)
.png){kind=icon}
.png){kind=icon}
分享.png){kind=icon}
收藏
摘要
En 中文
Olfactory receptors (ORs) belong to class A G-protein coupled receptors (GPCRs) and are activated by a variety of odorants. To date, there is no three-dimensional structure of an OR available. One of the major bottlenecks in obtaining purified protein for structural studies of ORs is their poor expression in heterologous cells. To design mutants that enhance expression and thereby enable protein purification, we first identified computable physical properties that recapitulate OR and class A GPCR expression and further conducted an iterative computational prediction-experimental test cycle and generated human OR mutants that express as high as biogenic amine receptors for which structures have been solved. In the process of developing the computational method to recapitulate the expression of ORs in membranes, we identified properties, such as amino acid sequence coevolution, and the strength of the interactions between intracellular loop 1 (ICL1) and the helix 8 region of ORs, to enhance their heterologous expression. We identified mutations that are directly located in these regions as well as other mutations not located in these regions but allosterically strengthen the ICL1-helix 8 enhance expression. These mutants also showed functional responses to known odorants. This method to enhance heterologous expression of mammalian ORs will facilitate high-throughput deorphanizationof ORs, and enable OR purification for biochemical and structural studies to understand odorant-OR interactions. SIGNIFICANCE Although the olfactory receptors (ORs) represent more than half of the class A GPCRs and are involved in many physiological processes, including the perception of smell, they remain one of the most unexplored class A GPCRsin terms of structure, ligand binding, and activation mechanism. The roadblock to studying the structure activity relationship in ORs is the inability to efficiently express ORs in heterologous cells. We have identified sequence and structure-based properties that influence their expression. Sequence coevolution and stability of intracellular loop regions recapitulate the expression level of 110 ORs in heterologous cells. Mutations in human ORs predicted to stabilize the intracellular loop regions showed enhanced expression similar to other class A GPCRs with structures.
Keyword:
INTRACELLULAR RETENTION
FUNCTIONAL EXPRESSION
ENDOPLASMIC-RETICULUM
RESIDUES
MUTATIONS
FAMILY
AI总结
对已上传原文的论文进行重点信息的提取，主要内容包括：简要概述、研究摘要、背景介绍、关键亮点、图文解析、展望与总结。
期刊
IF:
3.1
论文数:
5.0W
被引数:
4.4W
机构
引用论文
Molecular regulation of lysophosphatidic acid receptor 1 trafficking to the cell surface.png)
CELLULAR SIGNALLINGOAAI
CELLULAR SIGNALLINGIF3.7
OAAI分享收藏分享收藏分享收藏
查看更多内容