Gustin from human parotid saliva is carbonic anhydrase VI.
Gustin, a zinc-metalloprotein constituting about 3% of human parotid saliva protein was previously isolated and characterized as a single polypeptide chain of 37kDa with one mole of zinc tightly bound to the protein. It exhibited biological activity activating calmodulin dependent bovine brain cAMP phosphodiesterase and was decreased in saliva of patients with loss of taste in whom taste buds showed a specific pathological morphology. Determination of its primary structure by amino acid sequence revealed it was identical with carbonic anhydrase (CA) [EC 4.2.1.1] VI and had two N-linked glycosylation sites. Analysis by reverse phase HPLC and SDS-PAGE before and after deglycosylation confirmed a single peak with molecular weight of the purified protein being 37kDa, the deglycosylated protein, 33kDa. N-linked carbohydrate chains contained N-acetyl glucosamine, galactose, mannose, and fucose interior to di, tri and tetra sialyated termini. By isoelectric focusing five increasingly acidic pI values were determined consistent with addition of sialic acid as the terminal carbohydrate residue on the N-linked glycoforms of the protein. Gustin was found to exhibit CA activity but was inhibited by known CA inhibitors in a different manner than CA I or II. These findings, consistent with analysis of previous investigators, indicate that parotid saliva gustin is CA VI.
Clinical Neuroscience Branch, NIMH, NIH, Bethesda, Maryland, USA.
B J
BJ
Thatcher
A E
AE
Doherty
E
E
Orvisky
B M
BM
Martin
R I
RI
Henkin
D000595:Amino Acid Sequence; D000818:Animals; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010306:Parotid Gland / Q000378:metabolism*; D012463:Saliva / Q000378:metabolism*; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D017421:Sequence Analysis
D018076:DNA, Complementary; D012471:Salivary Proteins and Peptides; D002256:Carbonic Anhydrases; C477051:carbonic anhydrase VI