Protein complexes of the plant plasma membrane resolved by Blue Native PAGE
(2004) In Physiologia Plantarum 121(4). p.546-555- Abstract
- With the characterization of the total genomes of Arabidopsis thaliana and Oryza sativa, several putative plasma membrane components have been identified. However, a lack of knowledge at the protein level, especially for hydrophobic proteins, have hampered analyses of physiological changes. To address whether protein complexes may be present in the native membrane, we subjected plasma membranes isolated from Spinacia oleracea leaves to blue-native polyacrylamide gel electrophoresis (BN-PAGE). BN-PAGE is well established in the separation of functional membrane protein complexes from mitochondria and chloroplasts, but a resolved protein complex pattern from PM of eukaryotic cells has previously not been reported. Using this method, protein... (More)
- With the characterization of the total genomes of Arabidopsis thaliana and Oryza sativa, several putative plasma membrane components have been identified. However, a lack of knowledge at the protein level, especially for hydrophobic proteins, have hampered analyses of physiological changes. To address whether protein complexes may be present in the native membrane, we subjected plasma membranes isolated from Spinacia oleracea leaves to blue-native polyacrylamide gel electrophoresis (BN-PAGE). BN-PAGE is well established in the separation of functional membrane protein complexes from mitochondria and chloroplasts, but a resolved protein complex pattern from PM of eukaryotic cells has previously not been reported. Using this method, protein complexes from Spinacia oleracea PM could be efficiently solubilized and separated, including the highly hydrophobic aquaporin (apparent molecular mass 230 kDa), a putative tetramer of H+-ATPase, and several less abundant complexes with apparent masses around or above 750 kDa. After denaturation and separation of the complexes into their subunits in a second dimension (SDS-PAGE), several of the complexes were identified as hydrophobic membrane proteins. Large amounts of protein (up to 1 mg) can be resolved in each lane, which suggests that the method could be used to study also low-abundance protein complexes, e.g. under different physiological conditions. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/271079
- author
- Kjell, Jonas LU ; Rasmusson, Allan LU ; Larsson, H and Widell, Susanne LU
- organization
- publishing date
- 2004
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Physiologia Plantarum
- volume
- 121
- issue
- 4
- pages
- 546 - 555
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- wos:000223101300003
- scopus:4344627295
- ISSN
- 0031-9317
- DOI
- 10.1111/j.1399-3054.2004.00354.x
- language
- English
- LU publication?
- yes
- id
- 50089a1d-949d-45d5-a488-c7b51b9e95d7 (old id 271079)
- date added to LUP
- 2016-04-01 16:13:56
- date last changed
- 2024-01-11 03:57:03
@article{50089a1d-949d-45d5-a488-c7b51b9e95d7,
abstract = {{With the characterization of the total genomes of Arabidopsis thaliana and Oryza sativa, several putative plasma membrane components have been identified. However, a lack of knowledge at the protein level, especially for hydrophobic proteins, have hampered analyses of physiological changes. To address whether protein complexes may be present in the native membrane, we subjected plasma membranes isolated from Spinacia oleracea leaves to blue-native polyacrylamide gel electrophoresis (BN-PAGE). BN-PAGE is well established in the separation of functional membrane protein complexes from mitochondria and chloroplasts, but a resolved protein complex pattern from PM of eukaryotic cells has previously not been reported. Using this method, protein complexes from Spinacia oleracea PM could be efficiently solubilized and separated, including the highly hydrophobic aquaporin (apparent molecular mass 230 kDa), a putative tetramer of H+-ATPase, and several less abundant complexes with apparent masses around or above 750 kDa. After denaturation and separation of the complexes into their subunits in a second dimension (SDS-PAGE), several of the complexes were identified as hydrophobic membrane proteins. Large amounts of protein (up to 1 mg) can be resolved in each lane, which suggests that the method could be used to study also low-abundance protein complexes, e.g. under different physiological conditions.}},
author = {{Kjell, Jonas and Rasmusson, Allan and Larsson, H and Widell, Susanne}},
issn = {{0031-9317}},
language = {{eng}},
number = {{4}},
pages = {{546--555}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Physiologia Plantarum}},
title = {{Protein complexes of the plant plasma membrane resolved by Blue Native PAGE}},
url = {{http://dx.doi.org/10.1111/j.1399-3054.2004.00354.x}},
doi = {{10.1111/j.1399-3054.2004.00354.x}},
volume = {{121}},
year = {{2004}},
}