Characterizing molecular interactions in different bacteriorhodopsin assemblies by single-molecule force spectroscopy

被引:75
作者
Sapra, KT
Besir, S
Oesterhelt, D
Muller, DJ
机构
[1] Tech Univ Dresden, Ctr Biotechnol, D-01307 Dresden, Germany
[2] Max Planck Inst Mol Cell Biol & Genet, D-01307 Dresden, Germany
[3] Max Planck Inst Biochem, D-82152 Martinsried, Germany
关键词
atomic force microscopy; membrane protein assembly; single-molecule force spectroscopy; membrane protein oligomerization; membrane protein stability;
D O I
10.1016/j.jmb.2005.10.080
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
Using single-molecule force spectroscopy we characterized inter- and intramolecular interactions stabilizing structural segments of individual bacteriorhodopsin (BR) molecules assembled into trimers and dimers, and monomers. While the assembly of BR did not vary the location of these structural segments, their intrinsic stability could change up to 70% increasing from monomer to dimer to trimer. Since each stable structural segment established one unfolding barrier, we conclude that the locations of unfolding barriers were determined by intramolecular interactions but that their strengths were strongly influenced by intermolecular interactions. Subtracting the unfolding forces of the BR trimer from that of monomer allowed us to calculate the contribution of inter- and intramolecular interactions to the membrane protein stabilization. Statistical analyses showed that the unfolding pathways of differently assembled BR molecules did not differ in their appearance but in their population. This suggests that in our experiments the membrane protein assembly does not necessarily change the location of unfolding barriers within the protein, but certainly their strengths, and thus alters the probability of a protein to choose certain unfolding pathways. (c) 2005 Elsevier Ltd. All rights reserved.
引用
收藏
页码:640 / 650
页数:11
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