Human catechol-O-methyltransferase: Cloning and expression of the membrane-associated form

Journal Article · Fri Feb 15 04:00:00 EST 1991 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.88.4.1416· OSTI ID:6045530

Bertocci, B; Miggiano, V; Da Prada, M; Dembic, Z; Lahm, H W; Malherbe, P ^[1]

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1. F. Hoffmann-La Roche Ltd., Basel (Switzerland)

A cDNA clone for human catechol-O-methyltransferase was isolated from a human hepatoma cell line (Hep G2) cDNA library by hybridization screening with a porcine cDNA probe. The cDNA clone was sequenced and found to have an insert of 1226 nucleotides. The deduced primary structure of hCOMT is composed of 271 amino acid residues with the predicted molecular mass of 30 kDa. At its N terminus it has a hydrophobic segment of 21 amino acid residues that may be responsible for insertion of hCOMT into the endoplasmic reticulum membrane. The primary structure of hCOMT exhibits high homology to the porcine partial cDNA sequence (93%). The deduced amino acid sequence contains two tryptic peptide sequences (T-22, T-33) found in porcine liver catechol-O-methyltransferase (CEMT). The coding region of hCOMT cDNA was placed under the control of the cytomegalovirus promoter to transfect human kidney 293 cells. The recombinant hCOMT was shown by immunoblot analysis to be mainly associated with the membrane fraction. RNA blot analysis revealed one COMT mRNA transcript of 1.4 kilobases in Hep G2 poly(A){sup +} RNA.

• OSTI ID: 6045530
• 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. 88:4; ISSN PNASA; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINES
AMINO ACID SEQUENCE
ANIMAL CELLS
ANIMALS
AROMATICS
CARBON-GROUP TRANSFERASES
CATECHOLAMINES
CELL CONSTITUENTS
CELL MEMBRANES
DISEASES
DNA
DNA SEQUENCING
DOMESTIC ANIMALS
ENDOPLASMIC RETICULUM
ENZYMES
HEPATOMAS
HYDROXY COMPOUNDS
MAMMALS
MEMBRANES
METHYL TRANSFERASES
MOLECULAR STRUCTURE
NEOPLASMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PHENOLS
POLYPHENOLS
RECOMBINANT DNA
STRUCTURAL CHEMICAL ANALYSIS
SWINE
TRANSFERASES
TUMOR CELLS
VERTEBRATES