Expression, purification, crystallization and preliminary X-ray analysis of a nucleoside kinase from the hyperthermophile Methanocaldococcus jannaschii.
Methanocaldococcus jannaschii nucleoside kinase (MjNK) is an ATP-dependent non-allosteric phosphotransferase that shows high catalytic activity for guanosine, inosine and cytidine. MjNK is a member of the phosphofructokinase B family, but participates in the biosynthesis of nucleoside monophosphates rather than in glycolysis. MjNK was crystallized as the apoenzyme as well as in complex with an ATP analogue and Mg2+. The latter crystal form was also soaked with fructose-6-phosphate. Synchrotron-radiation data were collected to 1.70 A for the apoenzyme crystals and 1.93 A for the complex crystals. All crystals exhibit orthorhombic symmetry; however, the apoenzyme crystals contain one monomer per asymmetric unit whereas the complex crystals contain a dimer.
Arnfors
Linda
L
Center for Structural Biochemistry, Department of Biosciences at Novum, Karolinska Institute, S-141 57 Huddinge, Sweden.
Hansen
Thomas
T
Meining
Winfried
W
Schönheit
Peter
P
Ladenstein
Rudolf
R
D000255:Adenosine Triphosphate / Q000737:chemistry; D001051:Apoenzymes / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods; D019281:Dimerization; D004926:Escherichia coli / Q000235:genetics; D019605:Euryarchaeota / Q000201:enzymology*; D005636:Fructosephosphates / Q000737:chemistry; D008274:Magnesium / Q000737:chemistry; D025221:Phosphofructokinases / Q000737:chemistry; D010770:Phosphotransferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D014835:Volatilization; D014961:X-Ray Diffraction
D001051:Apoenzymes; D001426:Bacterial Proteins; D005636:Fructosephosphates; C027618:fructose-6-phosphate; D000255:Adenosine Triphosphate; D010770:Phosphotransferases; D025221:Phosphofructokinases; C021892:nucleoside phosphotransferase; D008274:Magnesium