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Expression and purification of rat glucose transporter 1 in Pichia pastoris

Venskutonytė, Raminta LU ; Elbing, Karin LU and Lindkvist-Petersson, Karin LU (2018) In Methods in Molecular Biology 1713.
Abstract

Large amounts of pure and homogenous protein are a prerequisite for several biochemical and biophysical analyses, and in particular if aiming at resolving the three-dimensional protein structure. Here we describe the production of the rat glucose transporter 1 (GLUT1), a membrane protein facilitating the transport of glucose in cells. The protein is recombinantly expressed in the yeast Pichia pastoris. It is easily maintained and large-scale protein production in shaker flasks, as commonly performed in academic research laboratories, results in relatively high yields of membrane protein. The purification protocol describes all steps needed to obtain a pure and homogenous GLUT1 protein solution, including cell growth, membrane isolation,... (More)

Large amounts of pure and homogenous protein are a prerequisite for several biochemical and biophysical analyses, and in particular if aiming at resolving the three-dimensional protein structure. Here we describe the production of the rat glucose transporter 1 (GLUT1), a membrane protein facilitating the transport of glucose in cells. The protein is recombinantly expressed in the yeast Pichia pastoris. It is easily maintained and large-scale protein production in shaker flasks, as commonly performed in academic research laboratories, results in relatively high yields of membrane protein. The purification protocol describes all steps needed to obtain a pure and homogenous GLUT1 protein solution, including cell growth, membrane isolation, and chromatographic purification methods.

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Please use this url to cite or link to this publication:
author
organization
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
keywords
Chromatography, Glucose transporter, GLUT1, Membrane protein, Pichia pastoris, Protein expression, Protein purification
in
Methods in Molecular Biology
volume
1713
pages
13 pages
publisher
Humana Press
external identifiers
  • scopus:85037571307
ISSN
1064-3745
DOI
10.1007/978-1-4939-7507-5_1
language
English
LU publication?
yes
id
1c28c46f-fe53-4595-9e5d-a4df391ebb95
date added to LUP
2017-12-21 08:32:25
date last changed
2018-01-07 12:29:30
@inbook{1c28c46f-fe53-4595-9e5d-a4df391ebb95,
  abstract     = {<p>Large amounts of pure and homogenous protein are a prerequisite for several biochemical and biophysical analyses, and in particular if aiming at resolving the three-dimensional protein structure. Here we describe the production of the rat glucose transporter 1 (GLUT1), a membrane protein facilitating the transport of glucose in cells. The protein is recombinantly expressed in the yeast Pichia pastoris. It is easily maintained and large-scale protein production in shaker flasks, as commonly performed in academic research laboratories, results in relatively high yields of membrane protein. The purification protocol describes all steps needed to obtain a pure and homogenous GLUT1 protein solution, including cell growth, membrane isolation, and chromatographic purification methods.</p>},
  author       = {Venskutonytė, Raminta and Elbing, Karin and Lindkvist-Petersson, Karin},
  issn         = {1064-3745},
  keyword      = {Chromatography,Glucose transporter,GLUT1,Membrane protein,Pichia pastoris,Protein expression,Protein purification},
  language     = {eng},
  pages        = {13},
  publisher    = {Humana Press},
  series       = {Methods in Molecular Biology},
  title        = {Expression and purification of rat glucose transporter 1 in Pichia pastoris},
  url          = {http://dx.doi.org/10.1007/978-1-4939-7507-5_1},
  volume       = {1713},
  year         = {2018},
}