Biosynthesis of human β2-adrenergic receptor in methylotrophic yeast Pichia pastoris and its purification View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2012-04

AUTHORS

A. S. Gerasimov, O. A. Zeinalov, M. A. El’darov, A. A. Shul’ga

ABSTRACT

Human β2-adrenergic receptor is among the most studied members of the family of G-protein coupled receptors. This receptor plays an important role in the work of vegetative nervous system and is a target for drugs used in heart and lung diseases. The crystal structure of the β2-adrenergic receptor was determined; however, its physiological role and molecular mechanisms of action are still not completely understood. In the present work, the pVR2ADRH vector which contains the gene encoding human β2-adrenergic receptor with a polyhistidine label at the C-terminus was obtained. The Pichia pastoris methylotrophic yeast GS115 strain cells were transformed by this vector. The yield of human β2-adrenergic receptor as a result of the heterologous expression in the P. pastoris cells was 20 mg/L of the culture. The β2-adrenergic receptor was isolated from the membrane fraction of yeasts and purified by means of a metal chelate affine and ion-exchange chromatography. Active β2-adrenergic receptor was isolated using a chromatography on the CL-4B alprenololsepharose. The final yield of the purified β2-adrenergic receptor was approximately 1 mg/L of the culture. A homogeneity of the β2-adrenergic receptor preparation was confirmed by a method of dynamic light scattering of a protein micellar solution. More... »

PAGES

279-286

Journal

TITLE

Molecular Biology

ISSUE

2

VOLUME

46

Author Affiliations

Identifiers

URI

http://scigraph.springernature.com/pub.10.1134/s0026893312020057

DOI

http://dx.doi.org/10.1134/s0026893312020057

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1050165704


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