Bacteriorhodopsin can be refolded from two independently stable transmembrane helices and the complementary five-helix fragment.
This paper describes experimental tests of the hypothesis that bacteriorhodopsin (BR) can fold by the association of independently stable transmembrane helices. Peptides containing the first and second helical segments of BR were chemically synthesized. These two peptides and the complementary five-helix fragment of BR were reconstituted in three separate populations of native-lipid vesicles which were then mixed and fused to allow the fragments to interact. After addition of retinal, absorption spectroscopy of the reconstituted BR and X-ray diffraction of two-dimensional crystals of this material showed that the native structure of BR was regenerated. The first two helices of BR can therefore be considered as independent folding domains, and covalent connections in the loops connecting the helices to each other and to the rest of the molecule are not essential for the appropriate association of the helices.
Kahn
T W
TW
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511.
Engelman
D M
DM
D000595:Amino Acid Sequence; D001436:Bacteriorhodopsins / Q000737:chemistry*; D003460:Crystallization; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012172:Retinaldehyde / Q000494:pharmacology; D013053:Spectrophotometry; D014961:X-Ray Diffraction
D001436:Bacteriorhodopsins; D012172:Retinaldehyde