1fj6
FRUCTOSE-1,6-BISPHOSPHATASE (MUTANT Y57W) PRODUCT/ZN COMPLEX (R-STATE)
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OverviewOverview
Wild-type porcine fructose-1,6-bisphosphatase (FBPase) has no tryptophan, residues. Hence, the mutation of Try57 to tryptophan places a unique, fluorescent probe in the structural element (loop 52-72) putatively, responsible for allosteric regulation of catalysis. On the basis of, steady-state kinetics, circular dichroism spectroscopy, and X-ray, crystallography, the mutation has little effect on the functional and, structural properties of the enzyme. Fluorescence intensity from the Trp57, mutant is maximal in the presence of divalent cations, fructose, 6-phosphate and orthophosphate, which together stabilize an R-state, conformation in which loop 52-72 is engaged with the active site. The, level of fluorescence emission decreases monotonically with increasing, levels of AMP, an allosteric inhibitor, which promotes the T-state, disengaged-loop conformation. The titration of various metal-product, complexes of the Trp57 mutant with fructose 2,6-bisphosphate (F26P(2)), causes similar decreases in fluorescence, suggesting that F26P(2) and AMP, individually induce similar conformational states in FBPase. Fluorescence, spectra, however, are sensitive to the type of divalent cation (Zn(2+), Mn(2+), or Mg(2+)) and suggest conformations in addition to the R-state, loop-engaged and T-state, loop-disengaged forms of FBPase. The work, presented here demonstrates the utility of fluorescence spectroscopy in, probing the conformational dynamics of FBPase.
About this StructureAbout this Structure
1FJ6 is a Single protein structure of sequence from Sus scrofa with F6P, ZN and PO4 as ligands. Active as Fructose-bisphosphatase, with EC number 3.1.3.11 Full crystallographic information is available from OCA.
ReferenceReference
Tryptophan fluorescence reveals the conformational state of a dynamic loop in recombinant porcine fructose-1,6-bisphosphatase., Nelson SW, Iancu CV, Choe JY, Honzatko RB, Fromm HJ, Biochemistry. 2000 Sep 12;39(36):11100-6. PMID:10998248
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