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Optimization of GPER-1 Expression an...

Coulter, Grace,

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Optimization of GPER-1 Expression and Purification for Single Particle Cryogenic-Electron Microscopy /

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Optimization of GPER-1 Expression and Purification for Single Particle Cryogenic-Electron Microscopy // Grace Coulter.
作者:	Coulter, Grace,
面頁冊數:	1 electronic resource (78 pages)
附註:	Source: Masters Abstracts International, Volume: 86-11.
Contained By:	Masters Abstracts International86-11.
標題:	Biochemistry. -
電子資源:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=32041409
ISBN:	9798315744276

Optimization of GPER-1 Expression and Purification for Single Particle Cryogenic-Electron Microscopy /
Coulter, Grace,

Optimization of GPER-1 Expression and Purification for Single Particle Cryogenic-Electron Microscopy /Grace Coulter. - 1 electronic resource (78 pages)

Source: Masters Abstracts International, Volume: 86-11.

G-protein coupled receptors (GPCRs) are a large and diverse family of membrane proteins that mediate cellular responses to extracellular signals via G-protein activation. The G-protein coupled estrogen receptor (GPER-1) has been proposed as a non-classical estrogen receptor and is widely expressed in human tissues and linked to various disease pathways, including breast cancer. Despite its therapeutic potential, the signaling mechanism of GPER-1 remains poorly understood, with conflicting reports about its interaction with estrogen and downstream signaling pathways. Recent advances in cryo-electron microscopy (cryo-EM) have enabled high-resolution structural studies of GPCR protein complexes, offering insights into receptor activation and G-protein specificity. A resolved cryo-EM structure of GPER-1 in complex with G-proteins could clarify the endogenous ligand interactions and signaling mechanism.This study focuses on optimizing the expression and purification of GPER-1 in complex with 17-Estradiol (E2), mini-Gq-protein, and the stabilizing antibody fragment scFv16 for single particle cryo-EM analysis. Insect cell expression systems were found to yield enough of GPER-1, but protein aggregation during purification posed a significant challenge. Improvements in detergent selection, buffer composition, particularly including 10% glycerol, and co-expression were identified as key factors to reduce protein aggregation. These enhancements led to improved complex assembly when analyzed with single-particle cryo-EM screening at New York Structural Biology Center (NYSBC).

English

ISBN: 9798315744276Subjects--Topical Terms:

518028
Biochemistry.
Subjects--Index Terms:

Cryo-electron microscopy

Optimization of GPER-1 Expression and Purification for Single Particle Cryogenic-Electron Microscopy /
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