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Isolation of inside-out thylakoid vesicles

Åkerlund, Hans-Erik LU (2000) In Aqueous Two-Phase Systems: Methods and Protocols p.167-175
Abstract
To understand the function of a biological membrane like that of chloroplast thylakoids, it is important to understand the arrangement of its different protein and lipid components. Preparations that have proven to be particularly suited for such studies are those consisting of membrane vesicles that are turned inside-out. Inside-out vesicles from the thylakoid membrane were first obtained from spinach chloroplasts by a combination of mechanical fragmentation and separation by aqueous two-phase partition (1,2). By the same or very similar procedures, inside-out thylakoid vesicles have now also been obtained from other plant sources such as pea (3), barley (4), mangrove (Avicennia marina) (5), lettuce (6), Euglena gracilis (7),... (More)
To understand the function of a biological membrane like that of chloroplast thylakoids, it is important to understand the arrangement of its different protein and lipid components. Preparations that have proven to be particularly suited for such studies are those consisting of membrane vesicles that are turned inside-out. Inside-out vesicles from the thylakoid membrane were first obtained from spinach chloroplasts by a combination of mechanical fragmentation and separation by aqueous two-phase partition (1,2). By the same or very similar procedures, inside-out thylakoid vesicles have now also been obtained from other plant sources such as pea (3), barley (4), mangrove (Avicennia marina) (5), lettuce (6), Euglena gracilis (7), cyanobacteria (8,9) and the photosynthetic bacteria Rhodopseudomonas viridis (10). Because the isolation procedure does not involve the use of detergents, the inside-out thylakoids have a preserved membrane structure and are ideally suited for structure-function studies. They have been used extensively in studies on thylakoid membrane topography (11-14) and for the identification of proteins associated with oxygen evolution (15,16). An important finding was that the inside-out vesicles are only formed from appressed (not exposed to stroma) thylakoid membranes, whereas right side-out vesicles derive from nonappressed (stroma exposed) membranes (17). As a result, the usefulness of inside-out vesicles could be extended to include studies on the lateral organization of the thylakoid membrane. The use of inside-out thylakoids in studies on structure and function of the thylakoid membrane has been reviewed by Andersson et al. (18). (Less)
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author
organization
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
in
Aqueous Two-Phase Systems: Methods and Protocols
editor
Hatti-Kaul, Rajni
pages
167 - 175
publisher
Human Press Inc, Totowa, NJ
ISBN
0-89603-541-7
DOI
10.1385/1-59259-028-4:167
language
English
LU publication?
yes
id
331edd93-cda1-4580-af40-77e98d8cd2a6 (old id 1474804)
date added to LUP
2009-09-29 18:10:17
date last changed
2016-04-16 07:21:57
@inbook{331edd93-cda1-4580-af40-77e98d8cd2a6,
  abstract     = {To understand the function of a biological membrane like that of chloroplast thylakoids, it is important to understand the arrangement of its different protein and lipid components. Preparations that have proven to be particularly suited for such studies are those consisting of membrane vesicles that are turned inside-out. Inside-out vesicles from the thylakoid membrane were first obtained from spinach chloroplasts by a combination of mechanical fragmentation and separation by aqueous two-phase partition (1,2). By the same or very similar procedures, inside-out thylakoid vesicles have now also been obtained from other plant sources such as pea (3), barley (4), mangrove (Avicennia marina) (5), lettuce (6), Euglena gracilis (7), cyanobacteria (8,9) and the photosynthetic bacteria Rhodopseudomonas viridis (10). Because the isolation procedure does not involve the use of detergents, the inside-out thylakoids have a preserved membrane structure and are ideally suited for structure-function studies. They have been used extensively in studies on thylakoid membrane topography (11-14) and for the identification of proteins associated with oxygen evolution (15,16). An important finding was that the inside-out vesicles are only formed from appressed (not exposed to stroma) thylakoid membranes, whereas right side-out vesicles derive from nonappressed (stroma exposed) membranes (17). As a result, the usefulness of inside-out vesicles could be extended to include studies on the lateral organization of the thylakoid membrane. The use of inside-out thylakoids in studies on structure and function of the thylakoid membrane has been reviewed by Andersson et al. (18).},
  author       = {Åkerlund, Hans-Erik},
  editor       = {Hatti-Kaul, Rajni},
  isbn         = {0-89603-541-7},
  language     = {eng},
  pages        = {167--175},
  publisher    = {Human Press Inc, Totowa, NJ},
  series       = {Aqueous Two-Phase Systems: Methods and Protocols},
  title        = {Isolation of inside-out thylakoid vesicles},
  url          = {http://dx.doi.org/10.1385/1-59259-028-4:167},
  year         = {2000},
}