Consequences of hydrophobic mismatch between lipids and melibiose permease on melibiose transport

被引:64
作者
Dumas, F
Tocanne, JF
Leblanc, G
Lebrun, MC
机构
[1] CNRS, Lab Pharmacol & Biol Struct, F-31077 Toulouse, France
[2] Univ Nice, Lab Physiol Membranes Cellulaires, LRC, CEA 16V, F-06238 Villefranche Sur Mer, France
[3] CNRS, ERS 1253, F-06238 Villefranche Sur Mer, France
关键词
D O I
10.1021/bi992634s
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The structural and functional consequences of a mismatch between the hydrophobic thickness rip Of a transmembrane protein and that d(L) of the supporting lipid bilayer were investigated using melibiose permease (MelB) from Escherichia coli reconstituted in a set of bis saturated and monounsaturated phosphatidylcholine species differing in acyl-chain length. Influence of MelB on the midpoint gel-to-liquid-phase transition temperature, T-m,, of the saturated lipids was investigated through fluorescence polarization experiments, with 1,6-diphenyl-1,3,5-hexatriene as the probe, for varying protein/lipid molar ratio. Diagrams in temperature versus MelB concentration showed positive or negative shifts in T-m with the short-chain lipids DiC12:0-PC and DiC14:0-PC or the long-chain lipids DiC16:0-PC and DiC18:OPC, respectively. Theoretical analysis of the data yielded a d(L) value of 3.0 +/- 0.1 nm for the protein, similar to the 3.02 nm estimated from hydropathy profiles. Influence of the acyl chain length on the carrier activity of MelB was investigated in the liquid phase, using the monounsaturated PCs. Binding of the sugar to the transporter showed no dependence on the acyl chain length. In contrast, counterflow and Delta Psi-driven experiments revealed strong dependence of melibiose transport on the lipid acyl chain length. Similar bell-shaped transport versus acyl chain length profiles were obtained, optimal activity being supported by diC16:l-PC. On account of a d(p) value of 2.65 nm for the lipid and of various local constraints which would all tend to elongate the acyl chains in contact with the protein, one can conclude that maximal activity was obtained when the hydrophobic thickness of the bilayer matched that of the protein.
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收藏
页码:4846 / 4854
页数:9
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