Isolation and Characterization of a Subunit Form of the Light-Harvesting Complex of Rhodospirillum rubrum

J. F. Miller, S. B. Hinchigeri, P. S. Parkes-Loach, J. R. Sprinkle, J. R. Riccobono, P. A. Loach*, P. M. Callahan

*Corresponding author for this work

Research output: Contribution to journalArticle

122 Scopus citations

Abstract

A new method is described for the isolation of subunits of the light-harvesting complex from Rhodospirillum rubrum (wild type and the G-9 mutant) in yields that approach 100%. The procedure involved treating membrane vesicles with ethylenediaminetetraacetic acid-Triton X-100 to remove components other than the light-harvesting complex and reaction center. In the preparation from wild-type cells, a benzene extraction was then employed to remove carotenoid and ubiquinone. The next step involved a careful addition of the detergent n-octyl ß-D-glucopyranoside, which resulted in a quantitative shift of the long-wavelength absorbance maximum from 873 to 820 nm. This latter complex was then separated from reaction centers by gel filtration on Sephadex G-100. The pigment-protein complex, now absorbing at 820 nm, contained two polypeptides of about 6-kilodalton molecular mass (referred to as a and ß) in a 1:1 ratio and two molecules of bacteriochlorophyll (BChl) for each αß pair. This complex is much smaller in size than the original complex absorbing at 873 nm but probably is an associated form such as α2ß2·4BChl or α3ß36·BChl. The 820-nm form could be completely shifted back to a form once again having a longer wavelength λmaxnear 873 nm by decreasing the octyl glucoside concentration. Thus, the complex absorbing at 820 nm appears to be a subunit form of the original 873-nm complex.

Original languageEnglish (US)
Pages (from-to)5055-5062
Number of pages8
JournalBiochemistry
Volume26
Issue number16
DOIs
StatePublished - 1987

ASJC Scopus subject areas

  • Biochemistry

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