Characterization of CA XIII, a novel member of the carbonic anhydrase isozyme family.
The carbonic anhydrase (CA) gene family has been reported to consist of at least 11 enzymatically active members and a few inactive homologous proteins. Recent analyses of human and mouse databases provided evidence that human and mouse genomes contain genes for still another novel CA isozyme hereby named CA XIII. In the present study, we modeled the structure of human CA XIII. This model revealed a globular molecule with high structural similarity to cytosolic isozymes, CA I, II, and III. Recombinant mouse CA XIII showed catalytic activity similar to those of mitochondrial CA V and cytosolic CA I, with k(cat)/K(m) of 4.3 x 10(7) m(-1) s(-1), and k(cat) of 8.3 x 10(4) s(-1). It is very susceptible to inhibition by sulfonamide and anionic inhibitors, with inhibition constants of 17 nm for acetazolamide, a clinically used sulfonamide, and of 0.25 microm, for cyanate, respectively. Using panels of cDNAs we evaluated human and mouse CA13 gene expression in a number of different tissues. In human tissues, positive signals were identified in the thymus, small intestine, spleen, prostate, ovary, colon, and testis. In mouse, positive tissues included the spleen, lung, kidney, heart, brain, skeletal muscle, and testis. We also investigated the cellular and subcellular localization of CA XIII in human and mouse tissues using an antibody raised against a polypeptide of 14 amino acids common for both human and mouse orthologues. Immunohistochemical staining showed a unique and widespread distribution pattern for CA XIII compared with the other cytosolic CA isozymes. In conclusion, the predicted amino acid sequence, structural model, distribution, and activity data suggest that CA XIII represents a novel enzyme, which may play important physiological roles in several organs.
Institute of Medical Technology, University of Tampere, Tampere, Finland.
Jonna
J
Lehtonen
Bairong
B
Shen
Mauno
M
Vihinen
Angela
A
Casini
Andrea
A
Scozzafava
Claudiu T
CT
Supuran
Anna-Kaisa
AK
Parkkila
Juha
J
Saarnio
Antti J
AJ
Kivelä
Abdul
A
Waheed
William S
WS
Sly
Seppo
S
Parkkila
D000595:Amino Acid Sequence; D000818:Animals; D002256:Carbonic Anhydrases / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D019143:Evolution, Molecular; D006801:Humans; D007527:Isoenzymes / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009928:Organ Specificity; D017421:Sequence Analysis
D007527:Isoenzymes; C483903:CA13 protein, human; D002256:Carbonic Anhydrases