Crystallization of acetate kinase from Methanosarcina thermophila and prediction of its fold.
The unique biochemical properties of acetate kinase present a classic conundrum in the study of the mechanism of enzyme-catalyzed phosphoryl transfer. Large, single crystals of acetate kinase from Methanosarcina thermophila were grown from a solution of ammonium sulfate in the presence of ATP. The crystals diffract to beyond 1.7 A resolution. Analysis of X-ray data from the crystals is consistent with a space group of C2 and unit cell dimensions a = 181 A, b = 67 A, c = 83 A, beta = 103 degrees. Diffraction data have been collected from the crystals at 110 and 277 K. Data collected at 277 K extend to lower resolution, but are more reproducible. The orientation of a noncrystallographic two-fold axis of symmetry has been determined. Based on an analysis of the predicted amino acid sequences of acetate kinase from several organisms, we hypothesize that acetate kinase is a member of the sugar kinase/actin/hsp70 structural family.
Buss
K A
KA
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907-1392, USA.
Ingram-Smith
C
C
Ferry
J G
JG
Sanders
D A
DA
Hasson
M S
MS
D000084:Acetate Kinase / Q000737:chemistry*; D000199:Actins / Q000737:chemistry; D000595:Amino Acid Sequence; D000645:Ammonium Sulfate; D017124:Conserved Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D019281:Dimerization; D004926:Escherichia coli / Q000201:enzymology; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry; D017020:Methanosarcina / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017510:Protein Folding; D016415:Sequence Alignment
D000199:Actins; D018840:HSP70 Heat-Shock Proteins; D000084:Acetate Kinase; D000645:Ammonium Sulfate