Human carbonic anhydrase IV: cDNA cloning, sequence comparison, and expression in COS cell membranes.

• DOI: 10.1073/pnas.89.4.1315
• Publication Date: 2006-05-31T12:16:50Z
• Authors: T Okuyama; S Sato; X L Zhu; A Waheed; W S Sly
• Keywords: 0303 health sciences; Base Sequence; Glycoside Hydrolases; Phosphoric Diester Hydrolases; Phosphatidylinositol Diacylglycerol-Lyase; Cell Membrane; Molecular Sequence Data; DNA; In Vitro Techniques; Transfection; 3. Good health; 03 medical and health sciences; Phosphoinositide Phospholipase C; Oligodeoxyribonucleotides; Solubility; Multigene Family; Chlorocebus aethiops; Animals; Humans; Amino Acid Sequence; Cloning, Molecular; Sequence Alignment; Carbonic Anhydrases

We have isolated a full-length cDNA for human carbonic anhydrase IV (CA IV) from lambda gt10 kidney library. The 1105-base-pair (bp) contains 47-bp 5' untranslated region, 936-bp open reading frame, and 122-bp 3' region. deduced amino acid sequence is colinear with the N-terminal of several tryptic peptides lung CA IV. It includes an 18-amino signal sequence, 260-amino region that shows 30-36% similarity 29-kDa cytoplasmic CAs I, II, III), additional 27-amino C-terminal ends in 21-amino hydrophobic domain. Of 17 "active site" residues are highly conserved other CAs, 16 also present Expression COS cells produced 35-kDa enzyme was membrane associated, resistant to inactivation by SDS, contained no carbohydrate, reacted on Western blots antiserum lung. Treatment membranes transfected phosphatidylinositol-specific phospholipase C released 20-30% expressed membranes, indicating at least anchored glycosyl-phosphatidylinositol linkage.