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  The role of lipids and salts in two-dimensional crystallization of the glycine–betaine transporter BetP from Corynebacterium glutamicum

Tsai, C.-J., Ejsing, C. S., Shevchenko, A., & Ziegler, C. (2007). The role of lipids and salts in two-dimensional crystallization of the glycine–betaine transporter BetP from Corynebacterium glutamicum. Journal of Structural Biology, 160(3), 275-286. doi:10.1016/j.jsb.2007.09.008.

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 Creators:
Tsai, Ching-Ju1, Author           
Ejsing, Christer S.2, Author
Shevchenko, Andrej2, Author
Ziegler, Christine1, Author           
Affiliations:
1Department of Structural Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068291              
2Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany, ou_persistent22              

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Free keywords: Membrane proteins; 2D crystallization; Lipid species; Mass spectroscopy; Multiple-precursor ion scanning; Electron microscopy
 Abstract: The osmoregulated and chill-sensitive glycine–betaine transporter (BetP) from Corynebacterium glutamicum was reconstituted into lipids to form two-dimensional (2D) crystals. The sensitivity of BetP partly bases on its interaction with lipids. Here we demonstrate that lipids and salts influence crystal morphology and crystallinity of a C-terminally truncated BetP. The salt type and concentration during crystallization determined whether crystals grew in the form of planar-tubes, sheets or vesicles, while the lipid type influenced crystal packing and order. Three different lipid preparations for 2D crystallization were compared. Only the use of lipids extracted from C. glutamicum cells led to the formation of large, well-ordered crystalline areas. To understand the lipid-derived influence on crystallinity, lipid extracts from different stages of the crystallization process were analyzed by quantitative multiple-precursor ion scanning mass spectroscopy (MS). Results show that BetP has a preference for fatty acid moieties 16:0–18:1, and that a phosphatidyl glycerol (PG) 16:0–18:1 rich preparation prevents formation of pseudo crystals.

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Language(s): eng - English
 Dates: 2007-09-062007-05-012007-09-122007-09-192007-12-01
 Publication Status: Published in print
 Pages: 12
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1016/j.jsb.2007.09.008
PMID: 17981051
 Degree: -

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Title: Journal of Structural Biology
  Abbreviation : J. Struct. Biol.
Source Genre: Journal
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Publ. Info: San Diego, CA : Elsevier
Pages: - Volume / Issue: 160 (3) Sequence Number: - Start / End Page: 275 - 286 Identifier: ISSN: 1047-8477
CoNE: https://pure.mpg.de/cone/journals/resource/954922650160