DELIPIDATION OF BACTERIORHODOPSIN AND RECONSTITUTION WITH EXOGENOUS PHOSPHOLIPID
DELIPIDATION OF BACTERIORHODOPSIN AND RECONSTITUTION WITH EXOGENOUS PHOSPHOLIPID
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DOI：
10.1073/pnas.77.1.323
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发表时间：
1980-01-01
期刊：
影响因子：
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通讯作者：
KHORANA, HG
中科院分区：
文献类型：
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作者：
HUANG, KS;BAYLEY, H;KHORANA, HG
用去污剂 Triton X-100 溶解来自盐杆菌的紫色膜，然后在脱氧胆酸盐溶液中进行凝胶过滤，得到细菌视紫红质，其 99% 以上不含内源脂质。脱脂的细菌视紫红质用外源磷脂重构以形成囊泡，该囊泡在光照下有效地易位质子。质子泵送的方向是从囊泡的外部到内部，表明囊泡中的细菌视紫红质的方向与细菌膜中的方向相反。通过从囊泡外部进行蛋白水解来切割蛋白质的羧基末端，证实了该方向。
Solubilization of the purple membrane from Halobacterium halobium with the detergent Triton X-100 followed by gel filtration in deoxycholate solution gave bacteriorhodopsin that was more than 99% free from endogenous lipid. The delipidated bacteriorhodopsin was reconstituted with exogenous phospholipids to form vesicles which on illumination efficiently translocated protons. The direction of proton pumping was from the outside to the interior of the vesicles, indicating that the orientation of bacteriorhodopsin in the vesicles was opposite to that in the bacterial membrane. This orientation was confirmed by cleavage of the carboxyl terminus of the protein by proteolysis from the outside of the vesicles.