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

Journal Article · Sat Feb 15 04:00:00 EST 1992 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.89.4.1315· OSTI ID:5560527

Okuyama, Torayuki; Sato, Seiji; Zhu, Xin Liang; Waheed, Abdul; Sly, W S ^[1]

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1. St. Louis Univ., MO (United States)

The authors have isolated a full-length cDNA for human carbonic anhydrase IV (CA IV) from a {lambda}gt10 human kidney cDNA library. The 1,105-base-pair (bp) cDNA contains a 47-bp 5{prime} untranslated region, a 936-bp open reading frame, and a 122-bp 3{prime} untranslated region. The deduced amino acid sequence is colinear with the N-terminal sequence and the sequence of several tryptic peptides of human lung CA IV. Expression of the cDNA in COS cells produced a 35-kDa enzyme that was membrane associated, resistant to inactivation by SDS, contained no carbohydrate, and reacted on Western blots with antiserum to the 35-kDa CA IV from human lung. Treatment of membranes from transfected COS cells with phosphatidylinositol-specific phospholipase C released 20-30% of the expressed enzyme from membranes, indicating that at least 20-30% of the expressed enzyme was anchored to membranes by a glycosyl-phosphatidylinositol linkage.

• OSTI ID: 5560527
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States) Vol. 89:4; ISSN PNASA; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBON-OXYGEN LYASES
CARBONIC ANHYDRASE
CELL CONSTITUENTS
CELL MEMBRANES
CLONING
DAYS LIVING RADIOISOTOPES
DNA
DNA HYBRIDIZATION
DNA SEQUENCING
DNA-CLONING
ENZYMES
HYBRIDIZATION
HYDRO-LYASES
ISOTOPES
KIDNEYS
LIGHT NUCLEI
LYASES
MEMBRANE PROTEINS
MEMBRANES
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PROTEINS
RADIOISOTOPES
RECOMBINANT DNA
STRUCTURAL CHEMICAL ANALYSIS