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PubMed · 8356065

Human mitochondrial carbonic anhydrase: cDNA cloning, expression, subcellular localization, and mapping to chromosome 16.

Y Nagao·J S Platero·A Waheed·W S Sly
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Abstract

A full-length cDNA clone encoding human mitochondrial carbonic anhydrase (CA), CA V, was isolated from a human liver cDNA library. The 1123-bp cDNA includes a 55-bp 5' untranslated region, a 915-bp open reading frame, and a 153-bp 3' untranslated region. Expression of the cDNA in COS cells produced active enzyme. The 34-kDa precursor and 30-kDa mature form of CA V were identified on Western blots of COS-cell homogenates by a CA V-specific antibody raised to a synthetic peptide corresponding to the C-terminal 17 aa of CA V. Both 34-kDa and 30-kDa bands were also present in mitochondria isolated from transfected COS cells, whereas only the 30-kDa band was present in mitochondria isolated from normal human liver. The N-terminal sequence determined directly on the 30-kDa soluble CA purified from transfected COS cells indicated that processing of the precursor to mature human CA V involves removal of a 38-aa mitochondrial leader sequence. The 267-aa sequence deduced for mature human CA V shows 30-49% similarity to amino acid sequences of previously characterized human CAs (CA I-CA VII) and 76% similarity to the corresponding amino acid sequence deduced from the mouse cDNA. PCR analysis of DNAs from human-rodent somatic cell hybrids localized the gene for CA V to human chromosome 16, the same chromosome to which CA VII has previously been mapped.

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Amino Acid SequenceAnimalsBase SequenceCarbonic AnhydrasesCell LineChromosome MappingChromosomes, Human, Pair 16Cloning, MolecularDNAHumansKineticsMitochondria, LiverMolecular Sequence DataOligodeoxyribonucleotidesPolymerase Chain ReactionRecombinant ProteinsSequence Homology, Amino AcidTransfectionComparative StudyJournal ArticleResearch Support, U.S. Gov't, P.H.S.

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Y Nagao, J S Platero, A Waheed, W S Sly. 1993-08-15. Human mitochondrial carbonic anhydrase: cDNA cloning, expression, subcellular localization, and mapping to chromosome 16.. https://doi.org/10.1073/pnas.90.16.7623

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