Crystallization and preliminary X-ray analysis of UMP/CMP-kinase from Dictyostelium discoideum with the specific bisubstrate inhibitor P1-(adenosine 5')-P5-(uridine 5')-pentaphosphate (UP5A).
UMP/CMP-kinase (UK) from the slime mold Dictyostelium discoideum has been purified to high homogeneity and co-crystallized with the bisubstrate inhibitor P1-(adenosine 5')-P5-(uridine 5')-pentaphosphate (UP5A). UP5A binds to UK with a dissociation constant (Kd) of 3 +/- 0.5 nM at 25 degrees C and pH 7.5. This is some 50-fold tighter than the binding of P1,P5-(diadenosine 5')-pentaphosphate (AP5A, Kd = 160 +/- 15 nM). AP5A is a bisubstrate inhibitor that is specific for adenylate kinase. The crystals have the symmetry of the tetragonal space group P4(1)2(1)2 or its enantiomorph P4(3)2(1)2. The unit cell dimensions are a = b = 78.5 A and c = 101.4 A. The crystals diffract to a Bragg spacing of 2.1 A.
Wiesmüller
L
L
Max-Planck-Institut für medizinische Forschung, Abteilung Biophysik, Heidelberg, Germany.
Scheffzek
K
K
Kliche
W
W
Goody
R S
RS
Wittinghofer
A
A
Reinstein
J
J
D000818:Animals; D001665:Binding Sites; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003460:Crystallization; D018360:Crystallography, X-Ray; D004023:Dictyostelium / Q000201:enzymology*; D015226:Dinucleoside Phosphates / Q000737:chemistry* / Q000378:metabolism; D004226:Dithioerythritol / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D009703:Nucleoside-Phosphate Kinase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D013050:Spectrometry, Fluorescence
D015226:Dinucleoside Phosphates; C093294:P(1)-(adenosine-5')-P(5)-(uridine-5')pentaphosphate; C012275:P(1),P(5)-di(adenosine-5'-)pentaphosphate; D004226:Dithioerythritol; C019979:cytidylate kinase; D009703:Nucleoside-Phosphate Kinase