Preliminary crystallographic studies of human mitochondrial NAD(P)(+)-dependent malic enzyme.
Human mitochondrial NAD(P)(+)-dependent malic enzyme was overexpressed in Escherichia coli and purified by anion-exchange, ATP affinity, and gel filtration chromatography. The protein was crystallized with the hanging-drop vapor diffusion method. Many different crystal forms were observed, five of which were characterized in some detail. A 2.5-A multiple-wavelength anomalous diffraction data set and a 2.1-A native data set were collected using synchrotron radiation on crystals containing selenomethionyl residues. These crystals belong to space group B2, with a = 204.4 A, b = 107.0 A, c = 59.2 A, and gamma = 101.9 degrees. Self-rotation functions demonstrated that the tetramer of this enzyme obeys 222 symmetry.
Bhargava
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G
Department of Biological Sciences, Columbia University, New York, New York, 10027, USA.
Mui
S
S
Pav
S
S
Wu
H
H
Loeber
G
G
Tong
L
L
D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D008291:Malate Dehydrogenase / Q000737:chemistry* / Q000648:ultrastructure; D008928:Mitochondria / Q000737:chemistry* / Q000201:enzymology*; D012645:Selenomethionine / Q000737:chemistry
D012645:Selenomethionine; D008291:Malate Dehydrogenase; C031917:malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+)