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Literature summary extracted from

  • Gee, C.L.; Nourse, A.; Hsin, A.Y.; Wu, Q.; Tyndall, J.D.; Grunewald, G.L.; McLeish, M.J.; Martin, J.L.
    Disulfide-linked dimers of human adrenaline synthesizing enzyme PNMT are catalytically active (2005), Biochim. Biophys. Acta, 1750, 82-92.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.28 expressed in Escherichia coli strain BL21(DE3)pLysS Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.1.28 hanging drop vapour diffusion method is used with 2 mircol drops on 3 M sealed over 500 mircol or 100 microl precipitant in 24-well or 96-well trays. In the absence of reducing agents crystals grow on protein concentrations of 30 to 40 mg/ml and appear in two or three days. The addition of DTT inhibits formation of crystals under the same condition. Reduced and oxidized glutathione are added to PEG/LiCl crystallisation conditions, crystals only grow in drops where the amount of oxidized glutathione is higher than reduced, the ratio of 1:20 (reduced glutathione/oxidized glutathione) gives the largest crystals. Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.28 C139A the C139A mutant separates into two forms on gel filtration, consistent with a monomer and dimer form, the equilibrium is significantly shifted in favour of the monomer. C139A mutant has similar kinetic constants as the wild type human phenylethanolamine N-methyltransferase Homo sapiens
2.1.1.28 C48A the C48A mutant separates into two forms on gel filtration, consistent with a monomer and dimer form, the equilibrium is significantly shifted in favour of the monomer. C48A mutant has similar kinetic constants as the wild type human phenylethanolamine N-methyltransferase. The monomer-dimer equilibrium in analytical ultracentrifugation for the dimer fractions of phenylethanolamine N-methyltransferase-His and C48A phenylethanolamine N-methyltransferase is 10fold lower than for the monomer fractions (Kd 35-64 microM and 305-551 microM, respectively). The relative Kd values show that C48A phenylethanolamine N-methyltransferase is less likely to form dimer than phenylethanolamine N-methyltransferase-His. Homo sapiens
2.1.1.28 C48A/C139A similar kinetic constants as the wild type human phenylethanolamine N-methyltransferase Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.28 0.0035
-
 S-adenosyl-L-methionine purified in the absence of reducing agent, dimer, His-tagged Homo sapiens
2.1.1.28 0.095
-
 phenylethanolamine purified in the absence of reducing agent, dimer, His-tagged Homo sapiens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.28 30000
-
 SDS-PAGE Homo sapiens
2.1.1.28 30280
-
 analytical ultracentrifugation, non tagged phenylethanolamine N-methyltransferase, monomer Homo sapiens
2.1.1.28 60550
-
 analytical ultracentrifugation, non tagged phenylethanolamine N-methyltransferase, dimer Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.28 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.28 6X-histidine tagged phenylethanolamine N-methyltransferase purified on a 10 ml column and with gel filtration. Non-tagged phenylethanolamine N-methyltransferase is purified with DEAE-Sepharose and gel filtration. Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.28 S-adenosyl-L-methionine + phenylethanolamine
-
 Homo sapiens S-adenosyl-L-homocysteine + N-methylphenylethanolamine
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.1.28 dimer gel filtration reveals the presence of two species at elution volumes consistent with monomeric and dimeric human phenylethanolamine N-methyltransferase. The more prominent peak corresponds to the dimer form. The amount of dimer can be reduced either by using a more elute concentration of the protein or by the addition of 0.5 mM DTT to the running buffer. SDS-PAGE can not distinguish between the two forms. Native PAGE clearly distinguishes between the two forms of human phenylethanolamine N-methyltransferase. Crystals from the dimer fraction grow faster. Monomer and dimer human phenylethanolamine N-methyltransferase have similar kinetic constatns. The monomer-dimer equilibruim in analytical ultracentrifugation for the dimer frations of phenylethanolamine N-methyltransferase-His and C48A phenylethanolamine N-methyltransferase is 10fold lower than for the monomer fractions (Kd 35-64 microM and 305-551 microM, respectively). Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
2.1.1.28 hPNMT
-
 Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.1.28 0.045
-
 phenylethanolamine purified in the absence of reducing agent, dimer, His-tagged Homo sapiens