title: The pH dependence of the allosteric response of human liver pyruvate kinase to fructose-1,6-bisphosphate, ATP, and alanine.
abstract: The allosteric regulation of human liver pyruvate kinase (hL-PYK) by fructose-1,6-bisphosphate (Fru-1,6-BP; activator), ATP (inhibitor) and alanine (Ala; inhibitor) was monitored over a pH range from 6.5 to 8.0 at 37 degrees C. As a function of increasing pH, hL-PYK's affinity for the substrate phosphoenolpyruvate (PEP), and for Fru-1,6-BP decreases, while affinities for ATP and alanine slightly increases. At pH 6.5, Fru-1,6-BP and ATP elicit only small allosteric impacts on PEP affinity. As pH increases, Fru-1,6-BP and ATP elicit greater allosteric responses, but the response to alanine is relatively constant. Since the magnitudes of the allosteric coupling for ATP and for alanine inhibition are different and the pH dependences of these magnitudes are not similar, these inhibitors likely elicit their responses using different molecular mechanisms. In addition, our results fail to support a general correlation between pH dependent changes in effector affinity and pH dependent changes in the corresponding allosteric response.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Kansas Medical Center, MS 3030, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA. afenton@kumc.edu
authors.forename: Aron W
authors.initials: AW
authors.lastname: Fenton
authors.forename: Myra
authors.initials: M
authors.lastname: Hutchinson
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000409:Alanine / Q000494:pharmacology*; D000494:Allosteric Regulation; D004926:Escherichia coli / Q000235:genetics; D005635:Fructosediphosphates / Q000494:pharmacology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000187:drug effects* / Q000201:enzymology; D011770:Pyruvate Kinase / Q000235:genetics / Q000378:metabolism*
chemical_list: D005635:Fructosediphosphates; D000255:Adenosine Triphosphate; D011770:Pyruvate Kinase; C029063:fructose-1,6-diphosphate; D000409:Alanine