Rabbit lung indolethylamine N-methyltransferase: cDNA and gene cloning and characterization

Michael A. Thompson, Richard M Weinshilboum

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Indolethylamine N-methyltransferase (INMT) catalyzes the N-methylation of tryptamine and structurally related compounds. This reaction has been studied because of its possible role in the in vivo synthesis of psychoactive compounds or neurotoxins and has been characterized biochemically in preparations of rabbit lung. Therefore, we set out to purify rabbit lung INMT, to clone and express its cDNA, and to clone and structurally characterize its gene as steps toward understanding the function and regulation of this enzyme. Rabbit lung INMT was purified and partial amino acid sequence was obtained. A polymerase chain reaction-based approach was then used to clone a rabbit lung INMT cDNA with a 792-base pair open reading frame that encoded a 263-amino acid protein with a predicted molecular mass of 29 kDa. When the cDNA was expressed in COS-1 cells, the encoded protein catalyzed the methylation of tryptamine and structurally related compounds, and was inhibited by two products of the reaction, S-adenosyl-L-homocysteine (AdoHcy) and N,N-dimethyltryptamine, as well as antimigraine drugs that are structurally related to N,N-dimethyltryptamine. Northern blot analysis demonstrated the presence of 2.0-kilobase mRNA species in rabbit lung, liver and, at lower levels, in brain. The cDNA was then used to clone the rabbit INMT gene. That gene had three exons and was structurally similar to the genes for nicotinamide N-methyltransferase and phenylethanolamine N- methyltransferase in several species. Cloning and expression of a rabbit lung INMT cDNA and cloning of the rabbit INMT gene represent important steps toward determination of the function and regulation of this mammalian methyltransferase enzyme.

Original languageEnglish (US)
Pages (from-to)34502-34510
Number of pages9
JournalJournal of Biological Chemistry
Volume273
Issue number51
DOIs
StatePublished - Dec 18 1998

Fingerprint

tryptamine N-methyltransferase
Cloning
Organism Cloning
Complementary DNA
Genes
Rabbits
Lung
N,N-Dimethyltryptamine
Clone Cells
Methylation
Nicotinamide N-Methyltransferase
Phenylethanolamine N-Methyltransferase
Amino Acids
Polymerase chain reaction
Neurotoxins
Methyltransferases
Molecular mass
Homocysteine
Enzymes
COS Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Rabbit lung indolethylamine N-methyltransferase : cDNA and gene cloning and characterization. / Thompson, Michael A.; Weinshilboum, Richard M.

In: Journal of Biological Chemistry, Vol. 273, No. 51, 18.12.1998, p. 34502-34510.

Research output: Contribution to journalArticle

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