The Use of HPLC for the Purification of the Q_в-Protein

Abstract

The Qв-protein is as a hydrophobic integral membrane protein firmly bound in the reaction center complex of photosystem II. A new method was developed to purify the SDS extracted protein using reversed-phase chromatography with two binary linear gradient systems. The identification of the Qв-protein was achieved by its rapidly labeling during photoassimilation of [35S]sulfate and by its reaction with the photoaffinity label azido-[14C]atrazine. Furtherm ore, antisera against the purified Qв-protein reacted with a single peak fraction, the second peak of the chrom atogram , which was identical with the labeled protein peak fraction.