The alpha and beta subunits of Cyanidium caldarium phycocyanin: properties and amino acid sequences at the amino terminus.
Phycocyanin was isolated and purified from the unicellular alga, Cyanidium caldarium. Subunits were prepared on a Bio-Rex-70 column developed stepwise with urea solutions (pH 1.9). The alpha subunit eluted in 8 M urea and the beta subunit eluted in 9 M urea. The alpha and beta subunits displayed absorption maxima at 660, 354, and 277 nm in 8 M and 9 M urea. The alpha:beta ratio of total absorbance under the 660-nm peak was 0.56 suggesting an alpha:beta phycocyanobilin chromophore ration of 1:2. On calibrated sodium dodecyl sulfate gels, the alphs subunit had an estimated molecular weight of 15,500 plus or minus 1100 and the beta subunit has an estimated molecular weight of 18,300 plus or minus 300. Minimum molecular weights based on one histidine residue per subunit were 16,300 for the alpha subunit and 18,750 for the beta subunit. Phycocyanin displayed a single visible absorption maximum at 625 nm and two positive circular dichroic bands at 632 and 610 nm. The alpha and beta subunits displayed single visible absorption maxima at 618 and 600 nm and single positive circular dichroic peaks at 620 and 585 nm, respectively. Two-dimensional maps of tryptic digests of the alpha and beta subunits revealed distinct patterns of peptides each of which was consistent with the lysine and arginine composition of these polypeptides. Maps of tryptic digests of phycocyanin contained 25 major peptides (a total of 27 lysine and arginine residues). Automated sequence analysis of separated subunits revealed a 70% homology within the first 27 residues at the amino terminus of the alpha and beta subunits of C. caldarium phycocyanin.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002850:Chromatography, Gel; D056890:Eukaryota / Q000032:analysis*; D046911:Macromolecular Substances; D010446:Peptide Fragments / Q000032:analysis; D010860:Pigments, Biological / Q000032:analysis; D010940:Plant Proteins / Q000032:analysis; D013053:Spectrophotometry
D000596:Amino Acids; D046911:Macromolecular Substances; D010446:Peptide Fragments; D010860:Pigments, Biological; D010940:Plant Proteins