Mouse histamine N-methyltransferase: cDNA cloning, expression, gene cloning and chromosomal localization

Liewei M Wang, L. Yan, C. McGuire, C. A. Kozak, M. Wang, U. J. Kim, M. Siciliano, Richard M Weinshilboum

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Objective: Histamine N-methyltransferase (HNMT) catalyzes the Nτ-methylation of histamine. We set out to clone a mouse liver HNMT cDNA and the mouse HNMT gene as steps toward characterizing molecular genetic mechanisms involved in the regulation of this important histamine-metabolizing enzyme. Design: A PCR-based strategy was used to clone both the mouse HNMT cDNA and the gene encoding that cDNA, Hnmt. The cDNA was used both to express recombinant mouse HNMT and to determine the chromosomal localization of Hnmt. Results: The mouse liver HNMT cDNA was 1657 bp in length with an 888 bp open reading frame (ORF) that encoded a 296 amino acid protein with a predicted Mr value of approximately 32.5 kDa. The amino acid sequence of the encoded protein was 84 % identical to that of human kidney HNMT. Mouse HNMT was expressed in COS-1 cells, and its apparent Km values for histamine and S-adenosyl-L-methionine (Ado-Met), the two cosubstrates for the reaction, were 5.3 and 5.8 μM, respectively. The mouse HNMT gene, Hnmt, spanned approximately 25 kb and had 7 exons. Its structure differed from that of the human gene primarily by the presence of an additional exon at the 5′-terminus. Hnmt mapped to mouse chromosome 2 in an area of conserved synteny to human chromosome 2q, the location of the human gene (2q22) on the basis of fluorescence in situ hybridization. Conclusions: Cloning and functional characterization of the mouse HNMT cDNA and gene will now make it possible to study in the mouse molecular genetic mechanisms involved the regulation of this important histamine-metabolizing enzyme.

Original languageEnglish (US)
Pages (from-to)300-308
Number of pages9
JournalInflammation Research
Volume50
Issue number6
StatePublished - 2001

Fingerprint

Histamine N-Methyltransferase
Cloning
Gene expression
Organism Cloning
Complementary DNA
Gene Expression
Genes
Histamine
Chromosomes
Liver
Molecular Biology
Exons
Clone Cells
Synteny
Amino Acids
S-Adenosylmethionine
Gene encoding
Methylation
Chromosomes, Human, Pair 2
COS Cells

Keywords

  • Histamine
  • Histamine N-methyltransferase
  • Methylation
  • Mouse cDNA
  • Mouse gene

ASJC Scopus subject areas

  • Toxicology
  • Pharmacology (medical)
  • Immunology
  • Cell Biology

Cite this

Mouse histamine N-methyltransferase : cDNA cloning, expression, gene cloning and chromosomal localization. / Wang, Liewei M; Yan, L.; McGuire, C.; Kozak, C. A.; Wang, M.; Kim, U. J.; Siciliano, M.; Weinshilboum, Richard M.

In: Inflammation Research, Vol. 50, No. 6, 2001, p. 300-308.

Research output: Contribution to journalArticle

Wang, LM, Yan, L, McGuire, C, Kozak, CA, Wang, M, Kim, UJ, Siciliano, M & Weinshilboum, RM 2001, 'Mouse histamine N-methyltransferase: cDNA cloning, expression, gene cloning and chromosomal localization', Inflammation Research, vol. 50, no. 6, pp. 300-308.
Wang, Liewei M ; Yan, L. ; McGuire, C. ; Kozak, C. A. ; Wang, M. ; Kim, U. J. ; Siciliano, M. ; Weinshilboum, Richard M. / Mouse histamine N-methyltransferase : cDNA cloning, expression, gene cloning and chromosomal localization. In: Inflammation Research. 2001 ; Vol. 50, No. 6. pp. 300-308.
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T2 - cDNA cloning, expression, gene cloning and chromosomal localization

AU - Wang, Liewei M

AU - Yan, L.

AU - McGuire, C.

AU - Kozak, C. A.

AU - Wang, M.

AU - Kim, U. J.

AU - Siciliano, M.

AU - Weinshilboum, Richard M

PY - 2001

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N2 - Objective: Histamine N-methyltransferase (HNMT) catalyzes the Nτ-methylation of histamine. We set out to clone a mouse liver HNMT cDNA and the mouse HNMT gene as steps toward characterizing molecular genetic mechanisms involved in the regulation of this important histamine-metabolizing enzyme. Design: A PCR-based strategy was used to clone both the mouse HNMT cDNA and the gene encoding that cDNA, Hnmt. The cDNA was used both to express recombinant mouse HNMT and to determine the chromosomal localization of Hnmt. Results: The mouse liver HNMT cDNA was 1657 bp in length with an 888 bp open reading frame (ORF) that encoded a 296 amino acid protein with a predicted Mr value of approximately 32.5 kDa. The amino acid sequence of the encoded protein was 84 % identical to that of human kidney HNMT. Mouse HNMT was expressed in COS-1 cells, and its apparent Km values for histamine and S-adenosyl-L-methionine (Ado-Met), the two cosubstrates for the reaction, were 5.3 and 5.8 μM, respectively. The mouse HNMT gene, Hnmt, spanned approximately 25 kb and had 7 exons. Its structure differed from that of the human gene primarily by the presence of an additional exon at the 5′-terminus. Hnmt mapped to mouse chromosome 2 in an area of conserved synteny to human chromosome 2q, the location of the human gene (2q22) on the basis of fluorescence in situ hybridization. Conclusions: Cloning and functional characterization of the mouse HNMT cDNA and gene will now make it possible to study in the mouse molecular genetic mechanisms involved the regulation of this important histamine-metabolizing enzyme.

AB - Objective: Histamine N-methyltransferase (HNMT) catalyzes the Nτ-methylation of histamine. We set out to clone a mouse liver HNMT cDNA and the mouse HNMT gene as steps toward characterizing molecular genetic mechanisms involved in the regulation of this important histamine-metabolizing enzyme. Design: A PCR-based strategy was used to clone both the mouse HNMT cDNA and the gene encoding that cDNA, Hnmt. The cDNA was used both to express recombinant mouse HNMT and to determine the chromosomal localization of Hnmt. Results: The mouse liver HNMT cDNA was 1657 bp in length with an 888 bp open reading frame (ORF) that encoded a 296 amino acid protein with a predicted Mr value of approximately 32.5 kDa. The amino acid sequence of the encoded protein was 84 % identical to that of human kidney HNMT. Mouse HNMT was expressed in COS-1 cells, and its apparent Km values for histamine and S-adenosyl-L-methionine (Ado-Met), the two cosubstrates for the reaction, were 5.3 and 5.8 μM, respectively. The mouse HNMT gene, Hnmt, spanned approximately 25 kb and had 7 exons. Its structure differed from that of the human gene primarily by the presence of an additional exon at the 5′-terminus. Hnmt mapped to mouse chromosome 2 in an area of conserved synteny to human chromosome 2q, the location of the human gene (2q22) on the basis of fluorescence in situ hybridization. Conclusions: Cloning and functional characterization of the mouse HNMT cDNA and gene will now make it possible to study in the mouse molecular genetic mechanisms involved the regulation of this important histamine-metabolizing enzyme.

KW - Histamine

KW - Histamine N-methyltransferase

KW - Methylation

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KW - Mouse gene

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