Literature summary for 2.1.1.28 extracted from

Wu, Q.; Gee, C.L.; Lin, F.; Tyndall, J.D.; Martin, J.L.; Grunewald, G.L.; McLeish, M.J.
Structural, mutagenic, and kinetic analysis of the binding of substrates and inhibitors of human phenylethanolamine N-methyltransferase (2005), J. Med. Chem., 48, 7243-7252.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli strain BL21(DE3)-pLysS	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
hanging drop vapour diffusion on 3 M tape with 1 microl protein/ligand mixture plus 1 microl precipitant over 100 microl precipitant (0.6-0.8 M NH4H2PO4, 0.1 M citrate pH 5.3-5.8). The human phenylethanolamine N-methyltransferase S-adenosyl-L-homocysteine SKF 64139 structure is solved by difference Fourier methods and refines at 2.4 A resolution. A hydrophilic inhibitor does not bind in a distinclty differnt orientation than a hydrophobic inhibitor.	Homo sapiens

Crystallization (Comment)

Organism

hanging drop vapour diffusion on 3 M tape with 1 microl protein/ligand mixture plus 1 microl precipitant over 100 microl precipitant (0.6-0.8 M NH4H2PO4, 0.1 M citrate pH 5.3-5.8). The human phenylethanolamine N-methyltransferase S-adenosyl-L-homocysteine SKF 64139 structure is solved by difference Fourier methods and refines at 2.4 A resolution. A hydrophilic inhibitor does not bind in a distinclty differnt orientation than a hydrophobic inhibitor.

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
D267A	higher Km and lower kcat values than wild type enzyme	Homo sapiens
E219A	higher Km and lower kcat values than wild type enzyme	Homo sapiens
K57A	higher Km and higher kcat values than wild type enzyme	Homo sapiens
V53A	higher Km and higher kcat values than wild type enzyme	Homo sapiens

Inhibitors

Inhibitors	Comment	Organism
2,3-dichloro-alpha-methylbenzylamine	-	Homo sapiens
3,4-dichloroamphetamine	-	Homo sapiens
7-iodo-1,2,3,4-tetrahydroisoquinoline	-	Homo sapiens
7-nitro-1,2,3,4-tetrahydroisoquinoline	-	Homo sapiens
LY134046	2,3,4,5-tetrahydro-1H-2-benzyzepine derivate with two chloro substituents on the aromatic ring. Is able to cross the blood-brain barrier.	Homo sapiens
SKF 29661	1,2,3,4-tetrahydroisoquinoline derivate	Homo sapiens
SKF 64139	1,2,3,4-tetrahydroisoquinoline derivate with two chloro substituents on the aromatic ring. Is able to cross the blood-brain barrier.	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.00055	-	3-trifluoromethylphenylethanolamine	wild type enzyme, assay containing S-adenosyl-L-methionine and 3-trifluoromethylphenylethanolamine	Homo sapiens
0.0012	-	S-adenosyl-L-methionine	wild type enzyme, assay containing S-adenosyl-L-methionine and 3-trifluoromethylphenylethanolamine	Homo sapiens
0.0015	-	anti-9-amino-6-(trifluoromethyl)benzonorbornene	wild type enzyme, assay containing S-adenosyl-L-methionine and anti-9-amino-6-(trifluoromethyl)benzonorbornene	Homo sapiens
0.002	-	S-adenosyl-L-methionine	wild type enzyme, assay containing S-adenosyl-L-methionine and anti-9-amino-6-(trifuoromethyl)benzonorbornene	Homo sapiens
0.0024	-	S-adenosyl-L-methionine	mutant D267, assay containing S-adenosyl-L-methionine and 3-trifluoromethylphenylethanolamine	Homo sapiens
0.0034	-	S-adenosyl-L-methionine	wild type enzyme, radiochemical assay containing S-adenosyl-L-methionine and phenylethanolamine	Homo sapiens
0.0051	-	S-adenosyl-L-methionine	mutant D267A, assay containing S-adenosyl-L-methionine and anti-9-amino-6-(trifluoromethyl)benzonorbornene	Homo sapiens
0.0071	-	S-adenosyl-L-methionine	mutant E219A and mutant D267N, radiochemical assay containing S-adenosyl-L-methionine and phenylethanolamine	Homo sapiens
0.0081	-	S-adenosyl-L-methionine	mutant D267A, radiochemical assay containing S-adenosyl-L-methionine and phenylethanolamine	Homo sapiens
0.0096	-	S-adenosyl-L-methionine	mutant V53A, radiochemical assay containing S-adenosyl-L-methionine and phenylethanolamine	Homo sapiens
0.0117	-	S-adenosyl-L-methionine	mutant K57A, radiochemical assay containing S-adenosyl-L-methionine and phenylethanolamine	Homo sapiens
0.025	-	anti-9-amino-6-(trifluoromethyl)benzonorbornene	mutant D267A, assay containing S-adenosyl-L-methionine and anti-9-amino-6-(trifluoromethyl)benzonorbornene	Homo sapiens
0.055	-	3-trifluoromethylphenylethanolamine	mutant D267, assay containing S-adenosyl-L-methionine and 3-trifluoromethylphenylethanolamine	Homo sapiens
0.1	-	phenylethanolamine	wild type enzyme, radiochemical assay containing S-adenosyl-L-methionine and phenylethanolamine	Homo sapiens
0.58	-	phenylethanolamine	mutant E219A, radiochemical assay containing S-adenosyl-L-methionine and phenylethanolamine	Homo sapiens
1.31	-	phenylethanolamine	mutant K57A, radiochemical assay containing S-adenosyl-L-methionine and phenylethanolamine	Homo sapiens
1.63	-	phenylethanolamine	mutant V53A, radiochemical assay containing S-adenosyl-L-methionine and phenylethanolamine	Homo sapiens
1.64	-	phenylethanolamine	mutant D267N, radiochemical assay containing S-adenosyl-L-methionine and phenylethanolamine	Homo sapiens
2.05	-	phenylethanolamine	mutant D267A, radiochemical assay containing S-adenosyl-L-methionine and phenylethanolamine	Homo sapiens

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P11086	expression plasmid pET17PNMT-his	-

Purification (Commentary)

Purification (Comment)	Organism
His-Select HC nickel affninity column and gel filtration	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
3-trifluoromethylphenylethanolamine + S-adenosyl-L-methionine	-	Homo sapiens	?	-	?
anti-9-amino-6-(trifluoromethyl)benzonorbornene + S-adenosyl-L-methionine	-	Homo sapiens	?	-	?
S-adenosyl-L-methionine + phenylethanolamine	-	Homo sapiens	S-adenosyl-L-homocysteine + N-methylphenylethanolamine	-	?

Synonyms

Synonyms	Comment	Organism
PNMT	-	Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.001	-	3-trifluoromethylphenylethanolamine	mutant D267A	Homo sapiens
0.0017	-	S-adenosyl-L-methionine	mutant D267N	Homo sapiens
0.0043	-	anti-9-amino-6-(trifluoromethyl)benzonorbornene	mutant D267A	Homo sapiens
0.0052	-	S-adenosyl-L-methionine	mutant D267A	Homo sapiens
0.0062	-	anti-9-amino-6-(trifluoromethyl)benzonorbornene	wild type enzyme	Homo sapiens
0.0083	-	3-trifluoromethylphenylethanolamine	wild type enzyme	Homo sapiens
0.0218	-	S-adenosyl-L-methionine	mutant E219A	Homo sapiens
0.0473	-	S-adenosyl-L-methionine	wild type enzyme	Homo sapiens
0.0567	-	S-adenosyl-L-methionine	mutant V53A	Homo sapiens
0.0658	-	S-adenosyl-L-methionine	mutant K57A	Homo sapiens

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism
0.00000155	-	SKF 64139	wild type enzyme, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.0000044	-	LY134046	wild type enzyme, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.000019	-	SKF 64139	mutant V53A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.0000225	-	LY134046	mutant V53A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.0000265	-	SKF 64139	mutant K57A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.00004	-	7-nitro-1,2,3,4-tetrahydroisoquinoline	wild type enzyme, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.000056	-	7-nitro-1,2,3,4-tetrahydroisoquinoline	binding of the inhibitor to the wild type human phenylethanolamine N-methyltransferase S-adenosyl-L-methionine complex	Homo sapiens
0.000057	-	SKF 29661	binding of the inhibitor to the wild type human phenylethanolamine N-methyltransferase S-adenosyl-L-methionine complex	Homo sapiens
0.000072	-	2,3-dichloro-alpha-methylbenzylamine	binding of the inhibitor to the wild type human phenylethanolamine N-methyltransferase S-adenosyl-L-methionine complex	Homo sapiens
0.000078	-	7-nitro-1,2,3,4-tetrahydroisoquinoline	wild type enzyme, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.0000906	-	LY134046	mutant K57A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.00012	-	SKF 29661	wild type enzyme, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.000121	-	2,3-dichloro-alpha-methylbenzylamine	wild type enzyme, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.00013	-	7-nitro-1,2,3,4-tetrahydroisoquinoline	mutant V53A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.00059	-	LY134046	mutant D267A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.00062	-	SKF 29661	mutant V53A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.00074	-	7-nitro-1,2,3,4-tetrahydroisoquinoline	mutant K57A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.000999	-	SKF 64139	mutant D267A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.00133	-	2,3-dichloro-alpha-methylbenzylamine	free wild type human phenylethanolamine N-methyltransferase	Homo sapiens
0.001375	-	SKF 64139	mutant E219A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.00227	-	3,4-dichloroamphetamine	binding of the inhibitor to the wild type human phenylethanolamine N-methyltransferase S-adenosyl-L-methionine complex	Homo sapiens
0.00236	-	7-nitro-1,2,3,4-tetrahydroisoquinoline	free wild type human phenylethanolamine N-methyltransferase	Homo sapiens
0.00284	-	SKF 29661	free wild type human phenylethanolamine N-methyltransferase	Homo sapiens
0.00312	-	LY134046	mutant E219A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.00364	-	3,4-dichloroamphetamine	wild type enzyme, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.0069	-	SKF 29661	mutant D57A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.0148	-	3,4-dichloroamphetamine	free wild type human phenylethanolamine N-methyltransferase	Homo sapiens
0.0179	-	7-nitro-1,2,3,4-tetrahydroisoquinoline	mutant E219A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.067	-	7-nitro-1,2,3,4-tetrahydroisoquinoline	mutant D267A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.189	-	SKF 29661	mutant E219A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens
0.321	-	SKF 29661	mutant D267A, inhibition assay containing a fixed concentration of S-adenosyl-L-methionine and variable concentrations of the inhibitor and phenylethanolamine	Homo sapiens