Expression and purification of rat glucose transporter 1 in Pichia pastoris
(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.
(Less)
- author
- Venskutonytė, Raminta LU ; Elbing, Karin LU and Lindkvist-Petersson, Karin LU
- organization
- publishing date
- 2018
- type
- Chapter in Book/Report/Conference proceeding
- publication status
- published
- subject
- keywords
- Chromatography, Glucose transporter, GLUT1, Membrane protein, Pichia pastoris, Protein expression, Protein purification
- host publication
- Methods in Molecular Biology
- series title
- Methods in Molecular Biology
- volume
- 1713
- pages
- 13 pages
- publisher
- Humana Press
- external identifiers
-
- scopus:85037571307
- pmid:29218513
- 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
- 2024-03-01 03:23:37
@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}},
booktitle = {{Methods in Molecular Biology}},
issn = {{1064-3745}},
keywords = {{Chromatography; Glucose transporter; GLUT1; Membrane protein; Pichia pastoris; Protein expression; Protein purification}},
language = {{eng}},
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}},
doi = {{10.1007/978-1-4939-7507-5_1}},
volume = {{1713}},
year = {{2018}},
}