Expression, purification and crystallization of human 5-lipoxygenase-activating protein with leukotriene-biosynthesis inhibitors.
The nuclear membrane protein 5-lipoxygenase-activating protein (FLAP) plays an essential role in leukotriene synthesis. Recombinant full-length human FLAP with a C-terminal hexahistidine tag has been expressed and purified from the cytoplasmic membrane of Escherichia coli. Diffraction-quality crystals of FLAP in complex with leukotriene-synthesis inhibitor MK-591 and with an iodinated analogue of MK-591 have been grown using the sitting-drop vapor-diffusion method. The crystals exhibit tetragonal symmetry (P42(1)2) and diffracted to a resolution limit of 4 A.
Xu
Shihua
S
Department of Medicinal Chemistry, Merck Research Laboratories, Rahway, NJ 07065, USA. shihua_xu@merck.com
McKeever
Brian M
BM
Wisniewski
Douglas
D
Miller
Douglas K
DK
Spencer
Robert H
RH
Chu
Lin
L
Ujjainwalla
Feroze
F
Yamin
Ting Ting
TT
Evans
Jilly F
JF
Becker
Joseph W
JW
Ferguson
Andrew D
AD
D058927:5-Lipoxygenase-Activating Proteins; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D006801:Humans; D015289:Leukotrienes / Q000096:biosynthesis*; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015394:Molecular Structure
D058927:5-Lipoxygenase-Activating Proteins; C548592:ALOX5AP protein, human; D002352:Carrier Proteins; D015289:Leukotrienes; D008565:Membrane Proteins