Improved membrane isolation in the purification of beta 2-adrenoceptors from transgenic Escherichia coli

Protein Expression and Purification
R A Hill, M N Sillence

Abstract

beta 2-Adrenoceptors (beta 2-AR) have been purified from many mammalian tissues. Unfortunately, other beta-AR subtypes expressed in the same cells are usually copurified, contaminating the preparation and interfering with subsequent investigations such as receptor characterization, ligand binding studies, immunoprecipitation, or development of anti-receptor antibodies. The advent of molecular biology techniques has facilitated the expression of beta 2-AR in cells in which no other similar molecules are present; thus, receptor purification has been simplified. beta 2-AR expressed in Escherichia coli provides a convenient source of receptor without the need for specialized culture facilities required for eukaryotic cells. The greater complexity of the gram-negative cell wall structure, however, complicates the purification of membrane-bound receptor from this source. In this report, we describe a reliable method for the partial purification of membrane-bound beta 2-AR from transgenic E. coli. Spheroplast formation followed by cell disruption and a carbonate wash procedure provided beta 2-AR bound to bacterial inner membrane in high yield.

References

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Citations

Aug 18, 2010·Se pu = Chinese journal of chromatography·Lihua XuLiujiao Bian
Apr 3, 2007·Biotechnology Progress·Emily C McCuskerAnne Skaja Robinson
Feb 28, 2009·Microbiology·Rhonda I HobbStuart A Thompson
Oct 19, 2010·Biochimica Et Biophysica Acta·Satita TapaneeyakornAnthony Watts
Sep 21, 2007·Molecular BioSystems·Filippo Mancia, Wayne A Hendrickson
Aug 20, 2019·Applied and Environmental Microbiology·Thomai P LazouChrysostomos I Dovas
Nov 5, 2019·Protein Expression and Purification·Daniel N WisemanAlan D Goddard

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