Crystallization of PNMT, the adrenaline-synthesizing enzyme, is critically dependent on a high protein concentration.
Phenylethanolamine N-methyltransferase, PNMT, utilizes the methylating cofactor S-adenosyl-L-methionine to catalyse the synthesis of adrenaline. Human PNMT has been crystallized in complex with an inhibitor and the cofactor product S-adenosyl-L-homocysteine using the hanging-drop technique with PEG 6000 and lithium chloride as precipitant. A critical requirement for crystallization was a high enzyme concentration (>90 mg ml(-1)) and cryocrystallography was used for high-quality data measurement. Diffraction data measured from a cryocooled crystal extend to a resolution of 2.3 A. Cryocooled crystals belong to space group P4(3)2(1)2 and have unit-cell parameters a = b = 94.3, c = 187.7 A.
Begun
Jake
J
Centre for Drug Design and Development and Special Research Centre for Functional and Applied Genomics, Institute for Molecular Bioscience, University of Queensland, Brisbane QLD 4072, Australia.
McLeish
Michael J
MJ
Caine
Joanne M
JM
Palant
Elka
E
Grunewald
Gary L
GL
Martin
Jennifer L
JL
D003460:Crystallization; D018360:Crystallography, X-Ray; D004837:Epinephrine / Q000378:metabolism; D006801:Humans; D018021:Lithium Chloride / Q000737:chemistry; D010625:Phenylethanolamine N-Methyltransferase / Q000737:chemistry* / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry; D011487:Protein Conformation; D012435:S-Adenosylhomocysteine / Q000737:chemistry / Q000378:metabolism
D011092:Polyethylene Glycols; D012435:S-Adenosylhomocysteine; D010625:Phenylethanolamine N-Methyltransferase; D018021:Lithium Chloride; D004837:Epinephrine