Expression of mouse carbonic anhydrase VII in E. coli and demonstration of its CO2 hydrase activity.
The alpha-carbonic anhydrase (alpha-CA) gene family in mammals encodes 10 CA or CA-like proteins (CA I-CA X). Although the gene for human CA VII has been cloned and characterized, the corresponding protein has not previously been purified, and hence, the CO2 hydrase activity of its product has not as yet been demonstrated. In this study, we have cloned the mouse CA VII cDNA in an E. coli, glutathione-S-transferase (GST) expression vector. The CO2 hydrase activity of the expressed protein is about 4% that of the high-activity CAII isozyme, demonstrating that this evolutionarily highly conserved protein is a catalytically active member of this CA gene family.
Lakkis
M M
MM
Department of Human Genetics, University of Michigan Medical School, Ann Arbor 48109, USA.
Bergenhem
N C
NC
Tashian
R E
RE
D000595:Amino Acid Sequence; D000818:Animals; D002256:Carbonic Anhydrases / Q000235:genetics* / Q000378:metabolism; D002384:Catalysis; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D010957:Plasmids
D018076:DNA, Complementary; D007527:Isoenzymes; D002256:Carbonic Anhydrases