title: Quercetin-3-rutinoside Inhibits Protein Disulfide Isomerase by Binding to Its b'x Domain.
abstract: Quercetin-3-rutinoside inhibits thrombus formation in a mouse model by inhibiting extracellular protein disulfide isomerase (PDI), an enzyme required for platelet thrombus formation and fibrin generation. Prior studies have identified PDI as a potential target for novel antithrombotic agents. Using a fluorescence enhancement-based assay and isothermal calorimetry, we show that quercetin-3-rutinoside directly binds to the b' domain of PDI with a 1:1 stoichiometry. The binding of quercetin-3-rutinoside to PDI induces a more compact conformation and restricts the conformational flexibility of PDI, as revealed by small angle x-ray scattering. The binding sites of quercetin-3-rutinoside to PDI were determined by studying its interaction with isolated fragments of PDI. Quercetin-3-rutinoside binds to the b'x domain of PDI. The infusion of the b'x fragment of PDI rescued thrombus formation that was inhibited by quercetin-3-rutinoside in a mouse thrombosis model. This b'x fragment does not possess reductase activity and, in the absence of quercetin-3-rutinoside, does not affect thrombus formation in vivo. The isolated b' domain of PDI has potential as an antidote to reverse the antithrombotic effect of quercetin-3-rutinoside by binding and neutralizing quercetin-3-rutinoside.
authors.affiliation: From the Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02115 and.
authors.forename: Lin
authors.initials: L
authors.lastname: Lin
authors.affiliation: From the Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02115 and.
authors.forename: Srila
authors.initials: S
authors.lastname: Gopal
authors.affiliation: From the Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02115 and.
authors.forename: Anish
authors.initials: A
authors.lastname: Sharda
authors.affiliation: From the Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02115 and.
authors.forename: Freda
authors.initials: F
authors.lastname: Passam
authors.affiliation: From the Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02115 and.
authors.forename: Sheryl R
authors.initials: SR
authors.lastname: Bowley
authors.affiliation: From the Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02115 and.
authors.forename: Jack
authors.initials: J
authors.lastname: Stopa
authors.affiliation: the Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou 350002, China.
authors.forename: Guangpu
authors.initials: G
authors.lastname: Xue
authors.affiliation: the Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou 350002, China.
authors.forename: Cai
authors.initials: C
authors.lastname: Yuan
authors.affiliation: From the Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02115 and.
authors.forename: Barbara C
authors.initials: BC
authors.lastname: Furie
authors.affiliation: From the Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02115 and.
authors.forename: Robert
authors.initials: R
authors.lastname: Flaumenhaft
authors.affiliation: From the Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02115 and.
authors.forename: Mingdong
authors.initials: M
authors.lastname: Huang
authors.affiliation: From the Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02115 and bfurie@bidmc.harvard.edu.
authors.forename: Bruce
authors.initials: B
authors.lastname: Furie
mesh_terms: D000818:Animals; D001665:Binding Sites; D002151:Calorimetry; D006801:Humans; D020128:Inhibitory Concentration 50; D051379:Mice; D008810:Mice, Inbred C57BL; D019704:Protein Disulfide-Isomerases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D012431:Rutin / Q000378:metabolism / Q000494:pharmacology*; D053838:Scattering, Small Angle; D013927:Thrombosis / Q000517:prevention & control; D014961:X-Ray Diffraction
chemical_list: D012431:Rutin; D019704:Protein Disulfide-Isomerases
keywords: PDI; coagulation factor; fibrin; isoquercetin; mouse; platelet; protein-disulfide isomerase; quercetin-3-rutinoside; thrombosis; thrombus formation