Lipid composition-dependent incorporation of multiple membrane proteins into liposomes

被引:23
作者
Daghastanli, KRP
Ferreira, RB
Thedei, G
Maggio, B
Ciancaglini, P
机构
[1] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Pret, Dept Quim, BR-14040901 Ribeirao Preto, Brazil
[2] Univ Sao Paulo, PCARP, Serv Bioterio Geral, Ribeirao Preto, Brazil
[3] Univ Uberaba, Inst Ciencias Biol & Saude, Uberaba, MG, Brazil
[4] Univ Nacl Cordoba, CIQUIBIC, Fac Ciencias Quim, Dept Quim Biol, RA-5000 Cordoba, Argentina
基金
巴西圣保罗研究基金会;
关键词
Pasteurella multocida; reconstitution; proteoliposome; bilayer thickness; hydrophobic matching; membrane proteins; lipid-protein co-solubilization;
D O I
10.1016/j.colsurfb.2004.03.015
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Membrane proteins from bacteria Pasteurella multocida were used as a model for studying its incorporation into liposomes. An important step to achieve efficient high yield protein incorporation in proteoliposomes is the study of the more suitable lipid composition. To this end, we compared the amount of total protein, reconstituted by co-solubilization methods, into liposomes of phospholipids with different polar head groups and acyl chain lengths. The liposomes and proteoliposomes were characterised by isopycnic centrifugation in sucrose gradient and by dynamic light scattering. Experimental and theoretical results were compared considering the effects exerted through the hydrocarbon chain length, volume, and optimal cross-sectional area of the phospholipid (combined in the geometrical critical packing parameter, lipid-protein matching), critical spontaneous radius of curvature of the bilayer vesicle, phase transition temperature of the lipid and ratio of lipid-protein molecules present in the vesicles. The highest incorporation of multiple proteins was found with dipalmitoylphosphatidylcholine (DPPC), reaching a yield of 93% compared to the lower relative amounts incorporated in proteoliposomes of the other lipids. The incorporation of multiple proteins induces a proportional enhancement of vesicular dimension, since DPPC-proteoliposomes have an average diameter of 1850 A, compared to the 1430 A for pure DPPC vesicles. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:127 / 137
页数:11
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