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Abstract:
The osmoregulated and chill-sensitive glycinebetaine 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 16:018:1, and that a phosphatidyl glycerol (PG) 16:018:1
rich preparation prevents formation of pseudo crystals.
2007 Elsevier Inc. All rights reserved.
