Research Topics
Species | P J FaySummaryAffiliation: University of Rochester Country: USA Publications
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Publications
Acidic residues C-terminal to the A2 domain facilitate thrombin-catalyzed activation of factor VIIIJennifer L Newell
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, New York 14642, USA
Biochemistry 47:8786-95. 2008..These results suggest that residues Glu720, Asp721, Glu724, and Asp725 likely constitute an exosite-interactive region in factor VIII facilitating cleavages for procofactor activation...
Mutating factor VIII: lessons from structure to functionPhilip J Fay
Department of Biochemistry, School of Medicine, University of Rochester, P O Box 712, 601 Elmwood Ave, Rochester, NY 14642, USA
Blood Rev 19:15-27. 2005..In this review, we document important and novel contributions following this line of investigation...
Contribution of factor VIII light-chain residues 2007-2016 to an activated protein C-interactive siteM Takeyama
Philip J Fay, Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, 601 Elmwood Avenue, Rochester, NY 14642, USA, Tel 1 585 275 6576, Fax 1 585 275 6007, E mail
Thromb Haemost 109:187-98. 2013..These results show a contribution of a number of residues within the 2007-2016 sequence, and in particular residues Met2010, Ser2011, and Leu2013 to an APC-interactive site...- The A1 and A2 subunits of factor VIIIa synergistically stimulate factor IXa catalytic activityP J Fay
Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA
J Biol Chem 274:15401-6. 1999.... - Cleavage of factor VIII heavy chain is required for the functional interaction of a2 subunit with factor IXAP J Fay
Department of Biochemistry and Biophysics and the Department of Medicine, University of Rochester School of Medicine, Rochester, New York 14642, USA
J Biol Chem 276:12434-9. 2001..These results indicate that HC cleavage by either thrombin or factor Xa is essential to expose the factor IXa-interactive site(s) in the A2 subunit required to modulate protease activity... - Human inhibitor antibodies specific for the factor VIII A2 domain disrupt the interaction between the subunit and factor IXaP J Fay
Department of Biochemistry, University of Rochester School of Medicine, Rochester, New York 14642, USA
J Biol Chem 274:29826-30. 1999..Furthermore, these results also suggest that factor VIII residues 484-509 contribute to a factor IXa-interactive site... - Factor VIIIa cofactor activity shows enhanced ionic strength sensitivity in the absence of phospholipidP J Fay
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, NY 14642, USA
Biochim Biophys Acta 1548:159-68. 2001.... - Activation of factor VIII and mechanisms of cofactor actionPhilip J Fay
Departments of Biochemistry and Biophysics and Medicine, PO Box 712, University of Rochester School of Medicine, Rochester, 601 Elmwood Avenue, Rochester, NY 14642, USA
Blood Rev 18:1-15. 2004..This effect is manifested in a dramatic increase in the catalytic rate constant, k(cat), by mechanisms that remain poorly understood... - The A2 subunit of factor VIIIa modulates the active site of factor IXaP J Fay
Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA
J Biol Chem 273:19049-54. 1998..These results indicate that the isolated A2 subunit modulates the active site of factor IXa and identifies a functional role for this subunit in factor VIIIa... - Factor VIII structure and functionPhilip J Fay
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, New York 14642, USA
Int J Hematol 83:103-8. 2006..Mechanisms for the down-regulation of factor Xase focus upon inactivation of the cofactor and include dissociation of the A2 subunit as well as activated protein C-catalyzed proteolysis... - Metal ion-independent association of factor VIII subunits and the roles of calcium and copper ions for cofactor activity and inter-subunit affinityH Wakabayashi
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, New York 14642, USA
Biochemistry 40:10293-300. 2001..Ca(2+) appears essential to promote the active conformation of factor VIII while Cu(2+) primarily enhances the intersubunit affinity... - Polymerization and RNase H activities of the reverse transcriptases from avian myeloblastosis, human immunodeficiency, and Moloney murine leukemia viruses are functionally uncoupledJ J DeStefano
Department of Biochemistry, University of Rochester, New York 14642
J Biol Chem 266:7423-31. 1991..Results demonstrate that the RNase H function is much less active than the polymerization function during processive DNA synthesis and that the activities are not strictly coupled... - Mutations associated with hemophilia A in the 558-565 loop of the factor VIIIa A2 subunit alter the catalytic activity of the factor Xase complexP Vincent Jenkins
Department of Biochemistry and Biophysics and the Department of Medicine, University of Rochester Medical Center, Rochester, NY 14642, USA
Blood 100:501-8. 2002..These results show that residues within the 558-565 loop are critical in modulating FIXa enzymatic activity but do not contribute significantly to the affinity of FVIIIa for FIXa... - Use of affinity-directed liquid chromatography-mass spectrometry to map the epitopes of a factor VIII inhibitor antibody fractionA E Griffiths
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, NY, USA
J Thromb Haemost 9:1534-40. 2011..Neutralizing factor (F) VIII antibodies develop in approximately 30% of individuals with hemophilia A and show specificity to multiple sites in the FVIII protein... - Requirements for the catalysis of strand transfer synthesis by retroviral DNA polymerasesR G Buiser
Department of Biochemistry, University of Rochester, New York 14642
J Biol Chem 266:13103-9. 1991..These results suggest that strand transfer synthesis relies on an unidentified functional activity present in RTs... - Factor IXa:factor VIIIa interaction. helix 330-338 of factor ixa interacts with residues 558-565 and spatially adjacent regions of the a2 subunit of factor VIIIaS P Bajaj
Department of Medicine, Saint Louis University School of Medicine, St Louis, Missouri 63104, USA
J Biol Chem 276:16302-9. 2001..These data support a conclusion that the helix 330 of factor IXa interacts with the A2 558-565 sequence. This information was used to model the interface between the IXa protease domain and the A2 subunit, which is also provided herein... - Residues 110-126 in the A1 domain of factor VIII contain a Ca2+ binding site required for cofactor activityHironao Wakabayashi
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, NY 14642, USA
J Biol Chem 279:12677-84. 2004.... - Clustered basic residues within segment 484-510 of the factor VIIIa A2 subunit contribute to the catalytic efficiency for factor Xa generationP V Jenkins
Department of Biochemistry, University of Rochester Medical Center, Rochester, NY, USA
J Thromb Haemost 2:452-8. 2004.... - Combining mutations of charged residues at the A2 domain interface enhances factor VIII stability over single point mutationsH Wakabayashi
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, NY 14642, USA
J Thromb Haemost 7:438-44. 2009..Blood, 112, 2761, 2008)... - Cleavage at Arg-1689 influences heavy chain cleavages during thrombin-catalyzed activation of factor VIIIJennifer L Newell
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, New York 14642, USA
J Biol Chem 284:11080-9. 2009..Overall, these results are consistent with a competition between heavy and light chains for thrombin exosite binding and subsequent proteolysis with binding of the former chain preferred... - Contribution of factor VIIIa A2 and A3-C1-C2 subunits to the affinity for factor IXa in factor XaseP Vincent Jenkins
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, New York 14642, USA
Biochemistry 43:5094-101. 2004..Thus both A2 and A3-C1-C2 subunits contribute to the affinity of FVIIIa for FIXa in the membrane-dependent FXase... - Cofactor activities of factor VIIIa and A2 subunit following cleavage of A1 subunit at Arg336Mary E Koszelak Rosenblum
Department of Biochemistry, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA
J Biol Chem 277:11664-9. 2002.... - Identification of a factor Xa-interactive site within residues 337-372 of the factor VIII heavy chainKeiji Nogami
Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA
J Biol Chem 279:15763-71. 2004.... - A Glu113Ala mutation within a factor VIII Ca2+-binding site enhances cofactor interactions in factor XaseHironao Wakabayashi
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, New York 14642, USA
Biochemistry 44:10298-304. 2005.... - Epitope mapping factor VIII A2 domain by affinity-directed mass spectrometry: residues 497-510 and 584-593 comprise a discontinuous epitope for the monoclonal antibody R8B12C Ansong
Department of Biochemistry and Biophysics, University of Rochester, School of Medicine, Rochester, NY 14642, USA
J Thromb Haemost 4:842-7. 2006..Furthermore, based upon blotting specificity, we speculate that residues 584-593 make a substantially greater contribution to the binding energy for this interaction... - Thrombin-catalyzed activation of factor VIII with His substituted for Arg372 at the P1 siteKeiji Nogami
Department of Biochemistry and Biophysics, PO Box 712, University of Rochester Medical Center, 601 Elmwood Ave, Rochester, NY 14642, USA
Blood 105:4362-8. 2005..These results indicate a markedly reduced rate of cleavage following substitution at the P(1)Arg, and this property likely reflects the severity of the hemophilia A phenotype... - Activated protein C-catalyzed inactivation of human factor VIII and factor VIIIa. Identification of cleavage sites and correlation of proteolysis with cofactor activityP J Fay
Department of Medicine, University of Rochester School of Medicine and Dentistry, New York 14642
J Biol Chem 266:20139-45. 1991..These results suggest a mechanism for activated protein C-catalyzed inactivation of factor VIII(alpha) involving both covalent alteration and fragment dissociation... - Interaction of the A1 subunit of factor VIIIa and the serine protease domain of factor X identified by zero-length cross-linkingK A Lapan
Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry Rochester, New York, USA
Thromb Haemost 80:418-22. 1998.... - A3 domain residue Glu1829 contributes to A2 subunit retention in factor VIIIaH Wakabayashi
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, NY 14642, USA
J Thromb Haemost 5:996-1001. 2007..A primary reason for the instability of FVIIIa is the tendency for the A2 subunit to dissociate from FVIIIa leading to an inactive cofactor and consequent loss of FXase activity... - The kissing hairpin sequence promotes recombination within the HIV-I 5' leader regionM Balakrishnan
Department of Biochemistry and Biophysics and Cancer Center, University of Rochester Medical Center, Rochester, New York 14642, USA
J Biol Chem 276:36482-92. 2001..Overall, these results suggest that template dimerization facilitated by the unique geometry of the DIS-promoted kissing interactions effectively promotes recombination within the HIV-I 5'-UTR... - Identification of residues contributing to A2 domain-dependent structural stability in factor VIII and factor VIIIaHironao Wakabayashi
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, New York 14642
J Biol Chem 283:11645-51. 2008..This observation is consistent with an altered conformation involving new inter-subunit interactions involving A2 domain following procofactor activation... - Factor VIII lacking the C2 domain retains cofactor activity in vitroHironao Wakabayashi
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, New York 14642, USA
J Biol Chem 285:25176-84. 2010..Together, these results indicate that FVIII domains other than C2, likely C1, make significant contributions to membrane-binding and membrane-dependent function... - Exosite-interactive regions in the A1 and A2 domains of factor VIII facilitate thrombin-catalyzed cleavage of heavy chainKeiji Nogami
Department of Biochemistry, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA
J Biol Chem 280:18476-87. 2005..These data suggest the acidic region comprising residues 389-394 in factor VIII A2 domain interacts with thrombin via its heparin-binding exosite and facilitates cleavage at Arg(740) during procofactor activation... - Relationship between plus strand DNA synthesis removal of downstream segments of RNA by human immunodeficiency virus, murine leukemia virus and avian myeloblastoma virus reverse transcriptasesG M Fuentes
Department of Microbiology, University of Rochester, School of Medicine and Dentistry, NY 14642, USA
Nucleic Acids Res 24:1719-26. 1996..This suggests that degradation of downstream pieces of RNA is not absolutely required before synthesis of the plus strand DNA. The implications of these findings for viral replication are discussed... - Proteolysis at Arg740 facilitates subsequent bond cleavages during thrombin-catalyzed activation of factor VIIIJennifer L Newell
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, New York 14642, USA
J Biol Chem 282:25367-75. 2007..Overall, these results support a model whereby cleavage of factor VIII by thrombin is an ordered pathway with cleavage at Arg740 facilitating cleavages at Arg372 and Arg1689, which result in procofactor activation... - Altered interactions between the A1 and A2 subunits of factor VIIIa following cleavage of A1 subunit by factor XaKeiji Nogami
Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA
J Biol Chem 278:1634-41. 2003..Furthermore, the C terminus of A1 contributes to the K(m) for factor X binding to factor Xase, and this parameter is critical for activity assessed in plasma-based assays... - Ca(2+) binding to both the heavy and light chains of factor VIII is required for cofactor activityHironao Wakabayashi
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, New York 14642, USA
Biochemistry 41:8485-92. 2002..These results suggest that factor VIII contains two Ca(2+) binding sites with different affinities and that active factor VIII can be reconstituted from HC and LC only when both chains are preactivated by Ca(2+)... - pH-dependent association of factor VIII chains: enhancement of affinity at physiological pH by Cu2+Hironao Wakabayashi
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, 601 Elmwood Ave, Rochester, NY 14642, USA
Biochim Biophys Acta 1764:1094-101. 2006..Furthermore, copper ion contributes to the maintenance of the heterodimer at physiologic pH by a mechanism consistent with bridging the two chains... - Identification of residues in the 558-loop of factor VIIIa A2 subunit that interact with factor IXaIndu Jagannathan
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, New York 14642, USA
J Biol Chem 284:32248-55. 2009..Furthermore, variants Y555A, V559A, D560A, G563A, I566A, and D569A showed >80% reduction in k(cat) for factor Xa generation. These results identify residues in the 558-loop critical to interaction with factor IXa in Xase... - Role of P1 residues Arg336 and Arg562 in the activated-Protein-C-catalysed inactivation of Factor VIIIaFatbardha Varfaj
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, NY 14642, USA
Biochem J 396:355-62. 2006..Thus, while P1 residues contribute to catalytic efficiency, residues removed from these sites make a primary contribution to the overall binding of APC to Factor VIIIa... - Mn2+ binding to factor VIII subunits and its effect on cofactor activityHironao Wakabayashi
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, New York, 14642, USA
Biochemistry 42:145-53. 2003..Thus, Mn2+appears to generate factor VIII cofactor activity by a similar mechanism as observed for Ca2+following its association at nonidentical sites on the protein... - Generation of enhanced stability factor VIII variants by replacement of charged residues at the A2 domain interfaceHironao Wakabayashi
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, NY, USA
Blood 112:2761-9. 2008..These results indicate that replacement of buried charged residues is an effective alternative to covalent modification in increasing factor VIII (VIIIa) stability... - Factor VIII A3 domain residues 1954-1961 represent an A1 domain-interactive siteCharles Ansong
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, New York 14642, USA
Biochemistry 44:8850-7. 2005..We propose that residues 1954-1961 of the A3 domain contribute to interactions with the A1 domain, facilitating their association in factor VIII... - Residues surrounding Arg336 and Arg562 contribute to the disparate rates of proteolysis of factor VIIIa catalyzed by activated protein CFatbardha Varfaj
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, NY 14642, USA
J Biol Chem 282:20264-72. 2007..Efficient cleavage at Arg(336) by APC is attributed to favorable P4-P3' residues at this site, whereas cleavage at Arg(562) can be accelerated following replacement with more optimal P4-P3' residues... - Mechanisms of factor Xa-catalyzed cleavage of the factor VIIIa A1 subunit resulting in cofactor inactivationKeiji Nogami
Department of Biochemistry, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA
J Biol Chem 278:16502-9. 2003..Furthermore, cleavage at Lys(36) appears to be selectively modulated by the C-terminal acidic region of A1, a region that may interact with factor Xa via its heparin-binding exosite... - Mechanisms of interactions of factor X and factor Xa with the acidic region in the factor VIII A1 domainKeiji Nogami
Department of Biochemistry, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA
J Biol Chem 279:33104-13. 2004..Furthermore, factor Xa but not factor X interacts with high affinity at this site via residues contained within the heparin-binding exosite of the proteinase... - Localization of a pH-dependent, A2 subunit-interactive surface within the factor VIIIa A1 subunitKeiji Nogami
Department of Biochemistry and Biophysics, University of Rochester Medical Center, P.O. Box 712, 601 Elmwood Avenue, Rochester, NY 14642, USA
Biochim Biophys Acta 1701:25-35. 2004..This interactive surface appears conformationally labile in the truncated A1 as judged by its apparent stabilization following association with A3-C1-C2... - Substrate requirements for secondary cleavage by HIV-1 reverse transcriptase RNase HMichele Wisniewski
Department of Biochemistry and Biophysics and the Cancer Center, University of Rochester, Rochester, New York 14642, USA
J Biol Chem 277:28400-10. 2002..However, NC protein greatly facilitated the secondary cut on the blunt-ended substrate, suggesting that NC compensates for the unfavorable substrate structure... - Mechanism of minus strand strong stop transfer in HIV-1 reverse transcriptionYan Chen
Department of Biochemistry and Biophysics, University of Rochester, New York 14642, USA
J Biol Chem 278:8006-17. 2003..The acceptor invades at gaps created by reverse transcriptase-RNase H in the donor-cDNA hybrid. The fragmented donor is eventually strand-displaced by the acceptor, completing the transfer... - Factor VIII A1 domain residues 97-105 represent a light chain-interactive siteCharles Ansong
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, New York 14642, USA
Biochemistry 45:13140-9. 2006..These results indicate that residues within segment 97-105 of the A1 domain interact with residues within the A3C1C2 domains of the light chain and contribute to the interface in the factor VIII heterodimer... - Exosite-dependent regulation of factor VIIIa by activated protein CChandrashekhara Manithody
Department of Biochemistry and Molecular Biology, St Louis University School of Medicine, MO 63104, USA
Blood 101:4802-7. 2003....
Research Grants
- STRUCTURE AND REGULATION OF HUMAN FACTOR VIIIPhilip Fay; Fiscal Year: 2005..Definition of these issues will yield valuable insights into the biochemistry of the native as well as dysfunctional factor VIII molecules, and provide information for the design of superior therapeutics. ..
- Factor Vllla interactions in the intrinsic factor XasePhilip Fay; Fiscal Year: 2007..Definition of these issues will yield valuable and fundamental insights into the biochemistry of the native as well as dysfunctional factor VIII molecules, and provide information for the design of superior therapeutics ..
- STRUCTURE AND REGULATION OF HUMAN FACTOR VIIIPhilip Fay; Fiscal Year: 2007....
- STRUCTURE AND REGULATION OF HUMAN FACTOR VIIIPhilip Fay; Fiscal Year: 2009....
- Factor VIIIa interactions in the intrinsic factor XasePhilip J Fay; Fiscal Year: 2010..In this application we will elucidate fine point structural details of inter-protein interactions that will define mechanisms for factor VIIIa cofactor function and the regulation of its activity in factor Xase. ..
- STRUCTURE AND REGULATION OF HUMAN FACTOR VIIIPhilip Fay; Fiscal Year: 1993..protection and the role of protein S in this reaction. Overall, these studies will offer insights into a coagulation protein central to hemostasis...
- STRUCTURE AND REGULATION OF HUMAN FACTOR VIIIPhilip Fay; Fiscal Year: 1992....
- STRUCTURE AND REGULATION OF HUMAN FACTOR VIIIPhilip Fay; Fiscal Year: 2001..Definition of these issues will yield valuable insights into the biochemistry of the native as well as dysfunctional factor VIII molecules, and provide information for the design of superior therapeutics. ..
- STRUCTURE AND REGULATION OF HUMAN FACTOR VIIIPhilip J Fay; Fiscal Year: 2010....