RRC ID 20799
Author Kamo T, Shimogawara K, Fukuzawa H, Muto S, Miyachi S.
Title Subunit constitution of carbonic anhydrase from Chlamydomonas reinhardtii.
Journal Eur J Biochem
Abstract Carbonic anhydrase purified from the cell surface of Chlamydomonas reinhardtii was inactivated by treatment with dithiothreitol. This treatment caused dissociation of the holoenzyme into 35-kDa (A) and 4-kDa (B) subunits as revealed by SDS/PAGE. The 35-kDa subunit was further separated into two components A1 (35 kDa) and A2 (36.5 kDa) by SDS/PAGE using a gradient gel. These two components have the same amino acid sequence up to at least the 10th amino acid from the N-terminus. The molecular masses were estimated at 76 kDa and 35 kDa for the holoenzyme and the large subunit, respectively, and the molar ratio of the former to the latter at 1:2, by using the techniques of low-angle laser light-scattering photometry and precision differential refractometry combined with gel-filtration HPLC. The molar ratio of the 35-kDa/4-kDa subunits was estimated at 1:1 the gel-filtration HPLC monitored with precision differential refractometry. Atomic-absorption spectrophotometry revealed that the holoenzyme contains two atoms of zinc. These results suggest that the holoenzyme is a heterotetramer composed of two large subunits (A1 and A2) and two small subunits (B).
Volume 192(2)
Pages 557-62
Published 1990-9-11
DOI 10.1111/j.1432-1033.1990.tb19261.x
PMID 2120056
MeSH Amino Acid Sequence Animals Carbonic Anhydrases / genetics* Carbonic Anhydrases / isolation & purification Cell Membrane / enzymology Chlamydomonas / enzymology* Chromatography, High Pressure Liquid Electrophoresis, Polyacrylamide Gel Humans Macromolecular Substances Molecular Sequence Data Molecular Weight Peptide Fragments / isolation & purification Peptide Mapping Sequence Homology, Nucleic Acid
Times Cited 44
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Algae NIES-2235