Reverse-Phase High-Performance Liquid Chromatography of Antenna Pigment- and Chlorosomal Proteins of Chloroflexus aurantiacus

Abstract

During the past few years, reverse-phase high-performance liquid chromatography (RP-HPLC) has emerged as one of the techniques best suited for the isolation and separation of low molecular weight proteins (Mr 5000) and polypeptide fragments generated by enzymic or chemical cleavage of proteins. (1, 2). The distinct advantages of RP-HPLC, speed, sensitivity, reproducibility and resolution, have contributed to great advances in protein structure analysis. Despite the success in isolating hydrophilic polypeptides of even considerable size (3), it was not until recently that RP-HPLC was employed as a final purification step in the isolation of hydrophobic membrane proteins. Examples are the separation of subunitsof chloroplast coupling factor (4), of mitochondrial cytochrome c-oxidase (5) and various structural envelope proteins of viral origin (sendai-, polio-, influenza-virus; 6, 7, 8).