Purification, crystallization and X-ray diffraction analysis of pavine N-methyltransferase from Thalictrum flavum.
A cDNA from the plant Thalictrum flavum encoding pavine N-methyltransferase, an enzyme belonging to a novel class of S-adenosylmethionine-dependent N-methyltransferases specific for benzylisoquinoline alkaloids, has been heterologously expressed in Escherichia coli. The enzyme was purified using affinity and gel-filtration chromatography and was crystallized in space group P2(1). The structure was solved at 2.0 A resolution using a xenon derivative and the single isomorphous replacement with anomalous scattering method.
Jain
Ankur
A
EMBL Hamburg Outstation, c/o DESY, Notkestrasse 85, D-22603 Hamburg, Germany.
Ziegler
Jörg
J
Liscombe
David K
DK
Facchini
Peter J
PJ
Tucker
Paul A
PA
Panjikar
Santosh
S
D003001:Cloning, Molecular; D003460:Crystallization; D008780:Methyltransferases / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D031949:Thalictrum / Q000201:enzymology*; D014961:X-Ray Diffraction
D010940:Plant Proteins; D011994:Recombinant Proteins; D008780:Methyltransferases