Preliminary X-ray analysis of a truncated form of recombinant fructose-2,6-bisphosphatase.
The bisphosphatase domain of rat liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase and a C-terminal 30 amino acid truncated form were expressed in high yield in Escherichia coli and purified to homogeneity. The separately expressed bisphosphatase domain and its C-terminal truncated form had kinetic properties similar to the bisphosphatase of the intact bifunctional enzyme, but their turnover numbers were fourfold higher. The truncated enzyme crystallized in space group P1 with two molecules per asymmetric unit. The determined cell dimensions are: a = 41.9 A, b = 43.5 A, c = 57.6 A, alpha = 95.2 degrees, beta = 99.3 degrees, and gamma = 106.2 degrees. These crystals diffract to 2.0 A resolution when exposed to synchrotron radiation and are suitable for crystallographic structure analysis.
Lee
Y H
YH
Department of Physiology and Biophysics, SUNY at Stony Brook 11794-8661.
Lin
K
K
Okar
D
D
Alfano
N L
NL
Sarma
R
R
Pflugrath
J W
JW
Pilkis
S J
SJ
D000818:Animals; D001483:Base Sequence; D018360:Crystallography, X-Ray; D017931:DNA Primers / Q000737:chemistry; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D010446:Peptide Fragments; D025481:Phosphofructokinase-2; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism / Q000648:ultrastructure*; D051381:Rats; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship
D017931:DNA Primers; D010446:Peptide Fragments; D011994:Recombinant Proteins; D025481:Phosphofructokinase-2; D010744:Phosphoric Monoester Hydrolases