Human secreted carbonic anhydrase: cDNA cloning, nucleotide sequence, and hybridization histochemistry
- The Howard Florey Institute of Experimental Physiology and Medicine, Parkville, Victoria (Australia)
Complementary DNA clones coding for the human secreted carbonic anhydrase isozyme (CAVI) have been isolated and their nucleotide sequences determined. These clones identify a 1.45-kb mRNA that is present in high levels in parotid submandibular salivary glands but absent in other tissues such as the sublingual gland, kidney, liver, and prostate gland. Hybridization histochemistry of human salivary glands shows mRNA for CA VI located in the acinar cells of these glands. The cDNA clones encode a protein of 308 amino acids that includes a 17 amino acid leader sequence typical of secreted proteins. The mature protein has 291 amino acids compared to 259 or 260 for the cytoplasmic isozymes, with most of the extra amino acids present as a carboxyl terminal extension. In comparison, sheep CA VI has a 45 amino acid extension. Overall the human CA VI protein has a sequence identity of 35 {percent} with human CA II, while residues involved in the active site of the enzymes have been conserved. The human and sheep secreted carbonic anhydrases have a sequence identity of 72 {percent}. This includes the two cysteine residues that are known to be involved in an intramolecular disulfide bond in the sheep CA VI. The enzyme is known to be glycosylated and three potential N-glycosylation sites (Asn-X-Thr/Ser) have been identified. Two of these are known to be glycosylated in sheep CA VI. Southern analysis of human DNA indicates that there is only one gene coding for CA VI.
- OSTI ID:
- 5735169
- Journal Information:
- Biochemistry; (USA), Vol. 30:2; ISSN 0006-2960
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
CARBONIC ANHYDRASE
SECRETION
RECOMBINANT DNA
DNA SEQUENCING
SALIVARY GLANDS
AUTORADIOGRAPHY
AMINO ACID SEQUENCE
CYTOCHEMISTRY
DNA HYBRIDIZATION
DNA-CLONING
ENDONUCLEASES
GENES
MAN
PHOSPHORUS 32
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMISTRY
BODY
CARBON-OXYGEN LYASES
CHEMISTRY
CLONING
DAYS LIVING RADIOISOTOPES
DNA
DNA-ASE
ENZYMES
ESTERASES
GLANDS
HYBRIDIZATION
HYDRO-LYASES
HYDROLASES
ISOTOPES
LIGHT NUCLEI
LYASES
MAMMALS
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PHOSPHODIESTERASES
PHOSPHORUS ISOTOPES
PRIMATES
RADIOISOTOPES
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
550601* - Medicine- Unsealed Radionuclides in Diagnostics