Molecular basis of the fructose-2,6-bisphosphatase reaction of PFKFB3: Transition state and the C-terminal function

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dc.contributor.authorM C Cavalier-
dc.contributor.authorSong-Gun Kim-
dc.contributor.authorD Neau-
dc.contributor.authorY H Lee-
dc.date.accessioned2017-04-19T09:28:56Z-
dc.date.available2017-04-19T09:28:56Z-
dc.date.issued2012-
dc.identifier.issn0887-3585-
dc.identifier.uri10.1002/prot.24015ko
dc.identifier.urihttps://oak.kribb.re.kr/handle/201005/10630-
dc.description.abstractThe molecular basis of fructose-2,6-bisphosphatase (F-2,6-P 2ase) of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB) was investigated using the crystal structures of the human inducible form (PFKFB3) in a phospho-enzyme intermediate state (PFKFB3-P F-6-P), in a transition state-analogous complex (PFKFB3 AlF 4), and in a complex with pyrophosphate (PFKFB3 PP i) at resolutions of 2.45, 2.2, and 2.3 A, respectively. Trapping the PFKFB3-P F-6-P intermediate was achieved by flash cooling the crystal during the reaction, and the PFKFB3 AlF 4 and PFKFB3 PP i complexes were obtained by soaking. The PFKFB3 AlF 4 and PFKFB3 PP i complexes resulted in removing F-6-P from the catalytic pocket. With these structures, the structures of the Michaelis complex and the transition state were extrapolated. For both the PFKFB3-P formation and break down, the phosphoryl donor and the acceptor are located within ∼5.1 A, and the pivotal point 2-P is on the same line, suggesting an "in-line" transfer with a direct inversion of phosphate configuration. The geometry suggests that NE2 of His253 undergoes a nucleophilic attack to form a covalent N-P bond, breaking the 2O-P bond in the substrate. The resulting high reactivity of the leaving group, 2O of F-6-P, is neutralized by a proton donated by Glu322. Negative charges on the equatorial oxygen of the transient bipyramidal phosphorane formed during the transfer are stabilized by Arg252, His387, and Asn259. The C-terminal domain (residues 440-446) was rearranged in PFKFB3 PP i, implying that this domain plays a critical role in binding of substrate to and release of product from the F-2,6-P 2ase catalytic pocket. These findings provide a new insight into the understanding of the phosphoryl transfer reaction.-
dc.publisherWiley-
dc.titleMolecular basis of the fructose-2,6-bisphosphatase reaction of PFKFB3: Transition state and the C-terminal function-
dc.title.alternativeMolecular basis of the fructose-2,6-bisphosphatase reaction of PFKFB3: Transition state and the C-terminal function-
dc.typeArticle-
dc.citation.titleProteins-Structure Function and Bioinformatics-
dc.citation.number4-
dc.citation.endPage1153-
dc.citation.startPage1143-
dc.citation.volume80-
dc.contributor.affiliatedAuthorSong-Gun Kim-
dc.contributor.alternativeNameCavalier-
dc.contributor.alternativeName김성건-
dc.contributor.alternativeNameNeau-
dc.contributor.alternativeName이용환-
dc.identifier.bibliographicCitationProteins-Structure Function and Bioinformatics, vol. 80, no. 4, pp. 1143-1153-
dc.identifier.doi10.1002/prot.24015-
dc.subject.keywordCatalysis-
dc.subject.keywordConformation-
dc.subject.keywordGlycolysis-
dc.subject.keywordTransition state-
dc.subject.keywordX-ray crystallography-
dc.subject.localCatalysis-
dc.subject.localconformation-
dc.subject.localConformation-
dc.subject.localglycolysis-
dc.subject.localGlycolysis-
dc.subject.localTransition state-
dc.subject.localX-ray crystallography-
dc.subject.localx-ray crystallography-
dc.description.journalClassY-
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Jeonbuk Branch Institute > Biological Resource Center > 1. Journal Articles
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