Kinetic analysis of interaction of BRCA1 tandem breast cancer c-terminal domains with phosphorylated peptides reveals two binding conformations.

Biochemistry
Yves NominéGeorges Mer

Abstract

Tandem breast cancer C-terminal (BRCT) domains, present in many DNA repair and cell cycle checkpoint signaling proteins, are phosphoprotein binding modules. The best-characterized tandem BRCT domains to date are from the protein BRCA1 (BRCA1-BRCT), an E3 ubiquitin ligase that has been linked to breast and ovarian cancer. While X-ray crystallography and NMR spectroscopy studies have uncovered the structural determinants of specificity of BRCA1-BRCT for phosphorylated peptides, a detailed kinetic and thermodynamic characterization of the interaction is also required to understand how structure and dynamics are connected and therefore better probe the mechanism of phosphopeptide recognition by BRCT domains. Through a global analysis of binding kinetics data obtained from surface plasmon resonance (SPR) and stopped-flow fluorescence spectroscopy, we show that the recognition mechanism is complex and best modeled by two equilibrium conformations of BRCA1-BRCT in the free state that both interact with a phosphopeptide, with dissociation constants ( K d) in the micromolar range. We show that the apparent global dissociation constant derived from this kinetic analysis is similar to the K d values measured using steady-state SPR, isothe...Continue Reading

References

Feb 14, 1971·Journal of Molecular Biology·B Lee, F M Richards
Sep 22, 1995·Journal of Molecular Biology·J Gómez, E Freire
Jan 1, 1995·Proteins·J Janin
Jul 9, 1996·Biochemistry·E R Guinto, E Di Cera
Jun 3, 1997·Biochemistry·A VindigniE Di Cera
May 19, 1998·Trends in Biochemical Sciences·D G Myszka, T A Morton
Sep 29, 1998·Methods in Enzymology·B M Baker, K P Murphy
Nov 11, 1999·Journal of Molecular Recognition : JMR·D G Myszka
Sep 27, 2001·Nature Structural Biology·P S BrzovicR E Klevit
Sep 27, 2001·Nature Structural Biology·R S WilliamsJ N Glover
Apr 23, 2002·Protein Science : a Publication of the Protein Society·Yasmina S N DayDavid G Myszka
Jul 5, 2002·Journal of Molecular Biology·C M S EkbladL S Itzhaki
Nov 13, 2002·The Journal of Biological Chemistry·R Scott Williams, J N Mark Glover
May 7, 2003·Proceedings of the National Academy of Sciences of the United States of America·Peter S BrzovicRachel Klevit
Oct 10, 2003·The Journal of Biological Chemistry·R Scott WilliamsJ N Mark Glover
Oct 25, 2003·Science·Isaac A MankeMichael B Yaffe
Oct 25, 2003·Science·Xiaochun YuJunjie Chen
May 11, 2004·Nature Structural & Molecular Biology·Julie A ClappertonStephen J Smerdon
May 11, 2004·Nature Structural & Molecular Biology·R Scott WilliamsJ N Mark Glover
Jul 10, 2004·Structure·Maria Victoria E BotuyanGeorges Mer
Mar 29, 2005·Biophysical Chemistry·Serapion PyrpassopoulosGeorge Nounesis
Mar 31, 2005·Annual Review of Physical Chemistry·Ninad V Prabhu, Kim A Sharp
Aug 17, 2005·Biochemistry·Ashok K VarmaJohn A A Ladias
Nov 22, 2005·Biophysical Journal·Binh NguyenW David Wilson
Apr 18, 2006·Proceedings of the National Academy of Sciences of the United States of America·Lauren K ElyJamie Rossjohn
May 16, 2006·Journal of Molecular Biology·Hind RayNicole Dalla Venezia
Jul 5, 2006·Genes & Development·Xiaochun YuJunjie Chen
Aug 10, 2006·Journal of Molecular Recognition : JMR·Donald J Winzor, Craig M Jackson
Feb 28, 1997·Biophysical Chemistry·G I MakhatadzeP L Privalov
Feb 20, 2007·PLoS Computational Biology·Rachel KarchinAndrej Sali
Mar 30, 2007·Proceedings of the National Academy of Sciences of the United States of America·Catherine M EakinRachel E Klevit

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Citations

Dec 4, 2010·The Journal of Biological Chemistry·Charles Chung Yun LeungJ N Mark Glover
Apr 15, 2014·Familial Cancer·Ida BiunnoRenato Mariani-Costantini
Dec 18, 2009·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Apr 14, 2012·Journal of Amino Acids·Markus Ritzefeld, Norbert Sewald
Feb 6, 2015·ACS Chemical Biology·E Railey WhiteMatthew C T Hartman

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