Separation of phycobiliprotein subunits by reverse-phase high-pressure liquid chromatography

✍ Scribed by Ronald V. Swanson; Alexander N. Glazer

• Publisher: Elsevier Science
• Year: 1990
• Tongue: English
• Weight: 917 KB
• Volume: 188
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(90)90609-d

✦ Synopsis

Baseline separation of subunits of diverse phycobiliproteins was achieved by a reverse-phase HPLC gradient method with a C4 large-pore column and a solvent system consisting of 0.1% trifluoroacetic acid (TFA) in water and 0.1% TFA in 2:1 (v/v) acetonitrile:isopropanol. The procedure was successfully applied to cyanobacterial allophycocyanin and C-phycocyanins, an unusual phycocyanin from a marine cyanobacterium, red algal B- and R-phycoerythrins, and a cryptomonad phycoerythrin. The subunit sizes in these proteins range from about 7.5 to 30 kDa. Sample recovery was in excess of 85% in all cases. On-line spectroscopic analysis with a multiple diode array detector allowed determination of the type and number of bilins carried by each subunit.