• Toxicological Research
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• v.6 no.1
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• pp.89-97
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• 1990

Neural regulation of phenylethanolamine N-meth-yltransferase (PNMT) was studied with reserpine as a neuronal agent in rat adrenal medulla. The enzyme activity assay and northern blot analysis were performed to determine whether the induction of PNMT activity after reserpine treatment was associated with elevation of mRNA coding for PNMT. The i.p. administration of reserpine (2.5 mg/kg) on alternate days fot 4 injections to rats brought about 30% increase of adrenal medullary PNMT activity and approximately 60% stimulation of the PNMT mRNA level in rat adrenal gland. A dose of 10 mg/kg of reserpine was chosen to perform optimum induction of PNMT activity in the rat adrenal gland based on the results of dose response curve of reserpine. Time course reserpine (10 mg/kg) effects on the rat adrenal medullary PNMT were as follows: 1. Peripheral PNMT activity reached maximum level after 7 days of drug treatment on alternate days. 2. Trans-synaptic stimulation by reserpine increased pretranslational activity of rat adrenal PNMT, but not translational activity. 3. Immunotitration of PNMT molecule after reserpine treatment indicated that reserpine produced an enzyme with greater antibody affinity than endogenous molecule in the rat adrenal gland.

• Bulletin of the Korean Chemical Society
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• v.32 no.2
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• pp.455-458
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• 2011

In previous report, with the aid of receptor-oriented pharmacophore-based in silico screening, we characterized three novel hPNMT inhibitors (YPN010, YPN016, and YPN017) and proposed that the hydrogen bonding interaction between inhibitors and side chain of Lys57 is very important to inhibitory activity of hPNMT. To confirm the importance of Lys57, mutant with substitution of Lys57 with Ala was cloned and binding study was performed for a K57A mutant of hPNMT using STD-NMR and fluorescence experiments. The binding constants for three novel inhibitors with mutant hPNMT were dramatically decreased compared to those with wild-type protein. K57A mutant-induced conversion of noradrenaline to adrenaline was suppressed about 95 % compared to wild-type hPNMT. Mutagenic analysis using a K57A mutant confirmed the importance of the Lys57 residue in binding of the inhibitor candidate to hPNMT as well as enzymatic activity of hPNMT, implying that these results are consistent with our binding model.

• Korean Journal of Cognitive Science
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• v.1 no.2
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• pp.361-376
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• 1989

Norepoine is N-methylated by the enzyme phenly ethanolamine N-metyltransferase(PNMT)to form epinephrine.this enzyme is larhly restructed to the adrenal medulla where epinephrine in mammalian brain where epinephrine function as a neurotransmitter.It seems clear that central epinephrine is involved in the regulation of cardiovacular function and in several forms of hypertension.However,information about the struture of mammalian epinephrine forming enzyme has been limited until now.But recently we isolate bovine and human PNMT cDNA clone using gtll expression library and sequcde total nucleotide composition.To obtain information about the structrue of the human and rat PNMT proteins and gones and to further define the extent of the evolutionary relationships among the PNMT molecules of these species human and rat genomic DNA clones to PNMT were sequentially isolated and characterized.

• The Microorganisms and Industry
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• v.16 no.1
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• pp.21-26
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• 1990

동물체의 신경전달물질은 acetylcholine을 비롯한 9종 이상의 물질이 알려져 있으며 이들 물질의 생체내에서의 기작과 생합성 과정에 관해서는 많이 알려져 있고 또한 이 방면의 연구도 활발하다. 그중에도 Joh와 Baetge등에 의하여 DBH와 PNMT 효소의 생산에 관여하는 mRNA가 면역침전법에 의하여 순수 분리되고 이를 주형으로 하여 DBH와 PNMT 효소 생산에 근본이 되는 유전자의 염기서열 및 유전자의 구조를 규명하는 작업이 진행되고 있으나 아직도 그 전체가 규명된 바는 없다 (Baetge등, 1981;1983;Joh등, 1983;1984). 그리하여 본인들은 상기의 연구자들로부터 PNMT 유전자인 cDNA를 M13mp18과 19의 vector phage에 재조합시키고 이 cDNA를 JM107rhk 109의 host bacteria에 도입하여 형질발현 실험을 통하여 확인하고자 하였다.

• Bulletin of the Korean Chemical Society
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• v.30 no.8
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• pp.1835-1838
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• 2009

Inhibition of human phenylethanolamine N-methyltransferase (hPNMT) has been proposed as a method for the treatment of several mental processes which related on adrenaline metabolism. We performed in silico screening to identify flavonoid inhibitors of hPNMT using automated docking method and selected 9 inhibitor candidates based on ligand score (LigScore) and binding free energy (${\Delta}G_{bind}$) estimation. Among 9 flavonoid candidates, 7 flavonoids belong to flavones while the rest of them belong to flavanone. All candidates have common chemical features; two hydrogen bond interactions with side chain of Lys75 and backbone carbonyl oxygen of Asn39, and two hydrophobic interactions. One hydrophobic site is formed by Val53, Leu262, and Met258 and the other is made up of Phe182, Ala186, Tyr222, and Val269. This study can be helpful to understand the structural features for inhibition of PNMT and showed flavonoids as promising inhibitor candidates for hPNMT.

• The Korean Journal of Pharmacology
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• v.24 no.1
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• pp.1-10
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• 1988

To obtain information about the structure of the human phenylethanolamin N-methyltransferase (PNMT) and to further define the extent of the evolutionary relationships among PNMT molecules of several spesies, a full length cDNA clone for bovine adrenal PNMT was used to screen a charon 4A genomic library. One phage was isolated and identified, which included the entire PNMT gene. The length of inserted genomic DNA was 13.1-Kilobase (Kb) containing two internal EcoRI sites. Construction of a restriction map and subsequent Southern and dot blot analysis with 5'-and3'-specific cDNA probes allowed the identification of exon-containing fragments. This is the first report of the cloning of gene for human epinephrine synthesizing enzyme.

• The Korean Journal of Pharmacology
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• v.32 no.2
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• pp.159-168
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• 1996

To determine the regulatory mechanism of phenylethanolamine N-methyltransferase (PNMT) in the adrenal gland and in central nervous system, we observed the change of enzyme activity and mRNA level of PNMT in the adrenal gland, the brain stem, and hypothalamus of rats, which were injected with two neuroleptic agents(reserpine and haloperidol ). Reserpine depleting catecholamines in presynaptic vesicle increased PNMT activities in the adrenal gland and the brain stem to 150% of the control in time-dependent manner, but not in the hypothalamus. Haloperidol blocking dopamine receptor decreased PNMT activities in the adrenal gland and the hypothalamus, but not in the brain stem. Thus, the results indicate that catecholamines inhibit synthesis of epinephrine in the brain stem and the adrenal gland, and that dopamine stimulates synthesis of epinephrine in the hypothalamus and the adrenal gland. In addition, since the change of mRNA levels were nearly in accordance with the change of activities, the transcriptional regulation of PNMT is considered the mechanism of the regulation of epinephrine neuron.

• Korean Journal of Veterinary Research
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• v.37 no.3
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• pp.489-497
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• 1997

The distributions characteristics of neurons displaying immunoreactivity to the catecholamine synthetic enzymes, tyrosine hydroxylase(TH), dopamine-${\beta}$-hydroxylase(DBH), and phenylethanolamine-N-methyltransferase(PNMT) were examined in the adjacent sections of the substantia nigra & ventral tegmentum of the Striped Field Mouse(Apodemus agrarius coreae). None of these cell groups displayed either DBH or PNMT immunoreactivity. Many TH-immunoreactive neurons were present in the substantia nigra & ventral tegmentum. The major dopaminergic cell(TH-positive, DBH- & PNMT-negative) group in the midbrain was present in the pars compacta of substantia nigra and adjacent ventral tegmentum. And smaller dopaminergic cell groups Were found in the pars reticulata of the substantia nigra and central liner nucleus.

• Development and Reproduction
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• v.6 no.2
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• pp.117-122
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• 2002

Dopamine(DA), norepinephrine(NE), and epinephrine(E) belong to a class of neurotransmitters known as catecholamine (CA) which are synthesized and secreted by mammalian brain and adrenal medulla. CA regulate several behavior patterns connected with breeding, and regulate GnRH-gonadotropin hormone axis' vitality between hypothalamus-pituitary gland linking with reproduction freeze. The present study examined effects of sex steroid hormone on the transcriptional activities of CA biosynthesis enzymes, tyrosine hydroxylase(TH), dopamine $\beta$ -hydroxylase(DBH), and phenylethaolamine-N-methyl transferase(PNMT). Mature female rats were ovariectomized(OVX) and implanted with 17 $\beta$-estradiol(E$_2$: 500 $\mu\textrm{g}$/ml) or sesame oil. Forty-eight hours after implantation all the animals were sacrificed. Total RNAs were extracted immediately and were applied to semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR). The expression level of TH was appeared by hypothalamus > SNc> adrenal medulla orders in OVX＋Oil group, and by SNc > hypothalamus) adrenal medulla orders in OVX+E$_2$ group. Treatment with E$_2$ significantly increased TH expression in SNc and adrenal medulla but in hypothalamus, the reduced TH expression was observed. The expression level of DBH was appeared by adrenal medulla > SNc > hypothalamus orders in OVX＋Oil group and in OVX＋E$_2$ group. Administration of E$_2$ significantly reduced DBH expression in SNc, and increased in adrenal medulla. Two cDNA products, large(PNMT1) and small(PMNTs) species of 110bp difference, were amplified in SNc and hypothalamus, but only PNMTs was observed in adenal medulla. The PNMTs expression level was in the order of adrenal medulla > hypothalamus > SNc in both OVX＋Oil and OVX＋E$_2$ group. The PNMTs expression in SNc and adrenal medulla was significantly increased byE$_2$. The present report demonstrated that estrogen effects on transcriptional activities for CA biosynthethic enzymes were tissue specific in adrenal medulla as well as different region of brain. These results suggest that it might be crucial relationship between the type of estrogen receptor and CA enzyme gene expression.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.114-114
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• 2002

포유동물에서 뇌와 부신에서 합성.분비되는 카테콜아민(Catecholamine, CA)계 신경전달물질인 dopamine(DA), norepinephrine(NE), epinephrine(E)은 체내 각종 생리현상의 조절에 필수적이며, 생식과 관련지어서는 시상하부-뇌하수체 간 GnRH-gonadotropin 호르몬 축의 활성을 조절하는 기능 외에도 번식과 관련된 여러 행동양식을 조절함이 잘 알려져 있다. 본 연구는 CA 생합성 효소들인 tyrosine hydroxylase(TH), dopamine beta-hydroxylase(DBH), phenylethanolamine-N-methyltransferase(PNMT)의 유전자 발현에 미치는 sex steroid의 영향을 조사하였다. 성숙한 암컷 횐쥐(SD strain)의 난소를 제거하고 1주 경과 후 vehicle(sesame oil; OVX+Oil 실험군) 또는 estradiol 17$\beta$(235ug/m1; OVX+E$_2$실험군)이 든 silastic capsule(길이 14mm; 내경 1.55mm; 외경 3.125mm)을 48 시간 동안 처리한 뒤 희생시켰다. 적출된 조직으로부터 RNA를 추출한 후 semi-quantitative RT-PCR을 시행하였다. (i) TH의 발현 정도는 OVX+Oil 군에서는 시상하부) substantia nigra(SNc)) 부신 순으로, OVX+E$_2$군에서는 SN글 부신) 시상하부 순으로 나타났다. TH 발현에 미치는 estradiol의 효과로 SNc과 부신에서는 유의한 증가를 보인데 비해 시상하부에서는 유의한 감소를 관찰하였다. (ii) DBH 발현 정도는 OVX+Oil군에서는 SNc> 부신> 시상 하부 순으로, OVX+E$_2$군에서는 부신＞ SNc＞ 시상하부 순이었다. DBH 발현에 미치는 estradiol의 효과로 SNc에서는 유의한 감소, 부신에서는 유의한 증가, 그리고 시상하부에서는 통계적 유의성은 없으나 감소하는 경향을 보였다. (iii) PNMT의 발현의 경우 SNc와 시상하부에서는 기보고된 바와 같이 alternative splicing에 의해 110bp 차이의 크고 작은 두 형태의 cDNA(PNMTI & PNMTs)가 증폭되었으나 부신에서는 작은 cDNA 만이 관찰되었다. PNMTs의 발현 정도는 OVX+Oil군과 OVX+E$_2$군에서 공히 부신＞ 시상하부＞ SNc 순이었고, PNMTI의 발현은 SNc가 시상하부 보다 다소 높은 경향이었으나 유의성은 없었다. PNMTs 발현에 미치는 estradiol의 효과로 SNc에서는 유의한 감소, 부신에서는 유의한 증가, 그리고 시상하부에서는 통계적 유의성은 없으나 증가하는 경향을 보였다. 본 연구에서는 CA 생합성 효소들의 유전자 발현의 조절에 미치는 estrogen 의 영향이 세포 기원이 neural crest cell인 부신 수질은 물론 뇌의 상이한 지역간에서도 조직특이적임을 관찰하였다. 이러한 결과는 각 조직에서의 estrogen 수용체 유형의 차이 혹은 작용 모드와 각 효소 유전자 발현 사이에 중요한 상관관계가 있음을 시사한다.