Unambiguous stereochemical course of rabbit liver fructose bisphosphatase hydrolysis.
The stereochemical course of rabbit liver fructose bisphosphatase (EC 3.1.3.11) was determined by hydrolyzing the substrate analogue (Sp)-[1-18O]fructose 1-phosphorothioate 6-phosphate in H(2)17O, incorporating the chiral, inorganic phosphorothioate product into adenosine 5'-O-(2-thiotriphosphate) (ATP beta S), and analyzing the isotopic distribution of 18O in ATP beta S by 31P NMR. The result indicates that the 1-phosphoryl group is transferred with inversion of configuration. A series of single-turnover experiments ruled out an acyl phosphate intermediate in the hydrolysis. Consequently, fructose bisphosphatase catalyzes the hydrolysis of fructose 1,6-bisphosphate via a direct transfer of the phosphoryl moiety to water.
Domanico
P L
PL
Rahil
J F
JF
Benkovic
S J
SJ
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives; D000818:Animals; D006597:Fructose-Bisphosphatase / Q000378:metabolism*; D005635:Fructosediphosphates; D006868:Hydrolysis; D066298:In Vitro Techniques; D008099:Liver / Q000201:enzymology*; D009682:Magnetic Resonance Spectroscopy; D011817:Rabbits; D013237:Stereoisomerism; D013379:Substrate Specificity; D013873:Thionucleotides
D005635:Fructosediphosphates; D013873:Thionucleotides; C022570:adenosine 5'-O-(2-thiotriphosphate); D000255:Adenosine Triphosphate; D006597:Fructose-Bisphosphatase; C029063:fructose-1,6-diphosphate