Accession Number:

ADA248455

Title:

Conformation of Membrane Proteins: Bacteriorhodopsin

Descriptive Note:

Final rept. 15 Apr 1989-14 Apr 1992

Corporate Author:

BRANDEIS UNIV WALTHAM MA

Personal Author(s):

Report Date:

1991-12-09

Pagination or Media Count:

11.0

Abstract:

One of the main goals of this research project was accomplished. A water soluble derivative of bacteriorhodopsin BR was synthesized. An activated species of methoxypolyethylene glycol MeOPEG was synthesized, yielding 2-0- methoxypolyethylene glycol-N-hydroxy succinimyl carbonate. MeO-PEG-SC. MeOPEG- SC was coupled with the purple membrane PM of Halobacterium hilobium to yield MeO-PEG-PM. This product was centrifuged and purified by washing with H20 76 conversion, dissolved in buffer with 5 sodium dodecylsulphate SDS, 1 1 and PAGE Electrophoresis performed. The separated MEO-PEG-BR band was recovered from the gel by electroelution, and the solution lyophilized. This material was dissolved in H20 and the SDS removed by passing the solution through an Extra- gel column Pierce and eluted with P04 buffer pH 7.0. The final solution was centrifuged at 200,000 g, yielding a clear water-soluble solution of MeO-PEGBR. The product could be reconstituted into miscells which were capable of proton pumping. The MeO-PEG-BR reconstituted into vesicles had physical-chemical properties identical with the original PM. The circular dichroism spectra, ultraviolet and visible spectrum, and fluorescence spectrum were identical to that of the PM. Proteins, Membranes, Conformation, Bacteriorhodopsin.

Subject Categories:

  • Medicine and Medical Research

Distribution Statement:

APPROVED FOR PUBLIC RELEASE