title: New interactions between tumor suppressor Fhit protein and a nonhydrolyzable analog of its AP4A substrate.
abstract: Fragile histidine triad protein (Fhit) is a protein which primarily hydrolyses dinucleoside polyphosphates. To investigate possible interactions between the protein and a substrate, we used a nonhydrolyzable phosphorothioate analog of Ap4A, containing 5-bromo-2'-deoxyuridine instead of one adenosine residue. Photocrosslinking, followed by LC-MS experiments, determined a complex in which the probe was covalently linked to the NDSIYEELQK peptide (residues 110-119). The peptide was located within the 'disordered' region, which is invisible in the known crystal structures of Fhit. This invisible and flexible part seems to play a role in the stabilization of the Fhit-substrate complex, which may be important for its tumor suppressor activity.
authors.affiliation: Department of Bioorganic Chemistry, Polish Academy of Sciences, Centre of Molecular and Macromolecular Studies, Lodz, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Krakowiak
authors.affiliation: Department of Bioorganic Chemistry, Polish Academy of Sciences, Centre of Molecular and Macromolecular Studies, Lodz, Poland.
authors.forename: Beata
authors.initials: B
authors.lastname: Kocoń-Rębowska
authors.affiliation: Department of Bioorganic Chemistry, Polish Academy of Sciences, Centre of Molecular and Macromolecular Studies, Lodz, Poland.
authors.forename: Rafał
authors.initials: R
authors.lastname: Dolot
authors.affiliation: Department of Bioorganic Chemistry, Polish Academy of Sciences, Centre of Molecular and Macromolecular Studies, Lodz, Poland.
authors.forename: Danuta
authors.initials: D
authors.lastname: Piotrzkowska
mesh_terms: D017766:Acid Anhydride Hydrolases / Q000378:metabolism*; D000595:Amino Acid Sequence; D001345:Autoradiography; D002853:Chromatography, Liquid; D003432:Cross-Linking Reagents / Q000378:metabolism; D015226:Dinucleoside Phosphates / Q000737:chemistry / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D006868:Hydrolysis; D008027:Light; D008958:Models, Molecular; D009363:Neoplasm Proteins / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity; D053719:Tandem Mass Spectrometry; D000099170:Bis(5'-adenosyl)triphosphatase
chemical_list: D017766:Acid Anhydride Hydrolases; D003432:Cross-Linking Reagents; D015226:Dinucleoside Phosphates; D009363:Neoplasm Proteins; D010455:Peptides; D011994:Recombinant Proteins; D000099170:Bis(5'-adenosyl)triphosphatase
keywords: Fhit; LC-MS; diadenosine polyphosphate; hydrolase; nucleoside phosphorothioate; photocrosslink