Abstract
S-Adenosylmethionine:histamine N-methyltransferase (EC 2.1.1.8.) (HMT) catalyzes the transfer of a methyl group from S-adenosyl-l-methionine to histamine resulting in the formation of tele-methylhistamine and S-adenosyl-l-homocysteine. In the present study, we purified HMT from bovine brain since this enzyme has been suggested to have some different kinetic properties from that from mouse or rat brain. The overall purification was about 8000-fold with a 2.9% yield. The specific activity of the final preparation was 517 nmol/min/mg protein. The pivotal steps in the purification consisted of ammonium sulfate precipitation, linear gradient DEAE-cellulose chromatography and elution with histamine and S-adenosylmethionine in DEAE cellulose chromatography. Bovine brain HMT could not be retained on the affinity columns using S-adenosylhomocysteine and histamine as ligands under the conditions tested. Molecular weight of the purified enzyme was determined to be 33,200 dalton by SDS-polyacrylamide gel electrophoresis. A pI value of 5.9 was obtained by isoelectric focusing. Amodiaquine and metoprine competitively inhibited the enzyme activity with respect to histamine with Ki values of 7.5 and 58 nM, respectively.
| Original language | English |
|---|---|
| Pages (from-to) | 135-141 |
| Number of pages | 7 |
| Journal | Neurochemistry International |
| Volume | 19 |
| Issue number | 1-2 |
| DOIs | |
| Publication status | Published - 1991 |
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ASJC Scopus subject areas
- Cell Biology
- Molecular Biology
- Cellular and Molecular Neuroscience
Cite this
Purification and partial characterization of histamine N-methyltransferase from bovine brain. / Nishibori, Masahiro; Oishi, Ryozo; Itoh, Yoshinori; Saeki, Kiyomi.
In: Neurochemistry International, Vol. 19, No. 1-2, 1991, p. 135-141.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Purification and partial characterization of histamine N-methyltransferase from bovine brain
AU - Nishibori, Masahiro
AU - Oishi, Ryozo
AU - Itoh, Yoshinori
AU - Saeki, Kiyomi
PY - 1991
Y1 - 1991
N2 - S-Adenosylmethionine:histamine N-methyltransferase (EC 2.1.1.8.) (HMT) catalyzes the transfer of a methyl group from S-adenosyl-l-methionine to histamine resulting in the formation of tele-methylhistamine and S-adenosyl-l-homocysteine. In the present study, we purified HMT from bovine brain since this enzyme has been suggested to have some different kinetic properties from that from mouse or rat brain. The overall purification was about 8000-fold with a 2.9% yield. The specific activity of the final preparation was 517 nmol/min/mg protein. The pivotal steps in the purification consisted of ammonium sulfate precipitation, linear gradient DEAE-cellulose chromatography and elution with histamine and S-adenosylmethionine in DEAE cellulose chromatography. Bovine brain HMT could not be retained on the affinity columns using S-adenosylhomocysteine and histamine as ligands under the conditions tested. Molecular weight of the purified enzyme was determined to be 33,200 dalton by SDS-polyacrylamide gel electrophoresis. A pI value of 5.9 was obtained by isoelectric focusing. Amodiaquine and metoprine competitively inhibited the enzyme activity with respect to histamine with Ki values of 7.5 and 58 nM, respectively.
AB - S-Adenosylmethionine:histamine N-methyltransferase (EC 2.1.1.8.) (HMT) catalyzes the transfer of a methyl group from S-adenosyl-l-methionine to histamine resulting in the formation of tele-methylhistamine and S-adenosyl-l-homocysteine. In the present study, we purified HMT from bovine brain since this enzyme has been suggested to have some different kinetic properties from that from mouse or rat brain. The overall purification was about 8000-fold with a 2.9% yield. The specific activity of the final preparation was 517 nmol/min/mg protein. The pivotal steps in the purification consisted of ammonium sulfate precipitation, linear gradient DEAE-cellulose chromatography and elution with histamine and S-adenosylmethionine in DEAE cellulose chromatography. Bovine brain HMT could not be retained on the affinity columns using S-adenosylhomocysteine and histamine as ligands under the conditions tested. Molecular weight of the purified enzyme was determined to be 33,200 dalton by SDS-polyacrylamide gel electrophoresis. A pI value of 5.9 was obtained by isoelectric focusing. Amodiaquine and metoprine competitively inhibited the enzyme activity with respect to histamine with Ki values of 7.5 and 58 nM, respectively.
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UR - http://www.scopus.com/inward/citedby.url?scp=0025891247&partnerID=8YFLogxK
U2 - 10.1016/0197-0186(91)90128-Z
DO - 10.1016/0197-0186(91)90128-Z
M3 - Article
AN - SCOPUS:0025891247
VL - 19
SP - 135
EP - 141
JO - Neurochemistry International
JF - Neurochemistry International
SN - 0197-0186
IS - 1-2
ER -