Mapping of the catalytic groove preferences of factor Xa reveals an inadequate selectivity for its macromolecule substrates.

The Journal of Biological Chemistry
Elsa P BianchiniB F Le Bonniec

Abstract

Factor Xa (FXa) hydrolyzes two peptide bonds in prothrombin having (Glu/Asp)-Gly-Arg-(Thr/Ile) for P(3)-P(2)-P(1)-P(1)' residues, but the exact preferences of its catalytic groove remain largely unknown. To investigate the specificity of FXa, we synthesized full sets of fluorescence-quenched substrates carrying all natural amino acids (except Cys) in P(3), P(2), P(1)', P(2)', and P(3)' and determined the k(cat)/K(m) values of cleavage. Contrary to expectation, glycine was not the "best" P(2) residue; peptide with phenylalanine was cleaved slightly faster. In fact, FXa had surprisingly limited preferences, barely more pronounced than trypsin; in P(2), the ratio of the k(cat)/K(m) values for the most favorable side chain over the least was 289 (12 with trypsin), but in P(1)', this ratio was only 30 (versus 80 with trypsin). This unexpected selectivity undoubtedly distinguished FXa from thrombin, which exhibited ratios higher than 19,000 in P(2) and P(1)'. Thus, with respect to the catalytic groove, FXa resembles a low efficiency trypsin rather than the highly selective thrombin. The rates of cleavage of the peptidyl substrates were virtually identical whether or not FXa was in complex with factor Va, suggesting that the cofactor ...Continue Reading

References

Jan 1, 1976·Methods in Enzymology·J Jesty, Y Nemerson
May 15, 1992·European Journal of Biochemistry·M T StubbsW Bode
Oct 29, 1991·Biochemistry·E W DavieW Kisiel
Jan 1, 1988·Annual Review of Biochemistry·K G MannS Krishnaswamy
Feb 15, 1988·European Journal of Biochemistry·S KawabataS Sakakibara
Jan 1, 1981·Methods in Enzymology·R LottenbergP L Coleman
Aug 5, 1993·Journal of Molecular Biology·K PadmanabhanW Kisiel
Oct 1, 1996·Proceedings of the National Academy of Sciences of the United States of America·Q D Dang, E Di Cera
Oct 7, 1997·Biochemistry·S Krishnaswamy, A Betz
Mar 21, 1998·The Journal of Biological Chemistry·R J BaughS Krishnaswamy
Jun 13, 1998·The Journal of Biological Chemistry·G L LongM Kalafatis
Jun 3, 1999·Biochimica Et Biophysica Acta·S GrahnH D Jakubke
Jan 8, 2000·The Journal of Biological Chemistry·P E MarqueB F Le Bonniec
May 11, 2000·Proceedings of the National Academy of Sciences of the United States of America·E CamererS R Coughlin
Jun 28, 2000·Proceedings of the National Academy of Sciences of the United States of America·J L HarrisC S Craik

❮ Previous
Next ❯

Citations

Oct 26, 2010·Analytical and Bioanalytical Chemistry·Vanessa Castro-LópezAnthony J Killard
Mar 14, 2012·The Journal of Biological Chemistry·Jennifer P DeAngelisPhilip J Fay
Apr 30, 2009·The Journal of Biological Chemistry·Peter G W Gettins, Steven T Olson
Jan 7, 2005·Journal of Thrombosis and Haemostasis : JTH·S Krishnaswamy
Aug 19, 2005·Journal of Bioinformatics and Computational Biology·Sarah E BoydMaria Garcia de la Banda
Mar 3, 2012·PloS One·Maike GallwitzLars Hellman
Dec 13, 2006·Current Opinion in Chemical Biology·Scott L Diamond
Mar 30, 2007·European Journal of Haematology·Sepideh AkhavanJacqueline Tapon-Bretaudière
Aug 23, 2006·Chemical Biology & Drug Design·Karen M BromfieldRobert N Pike
May 1, 2012·Journal of Peptide Science : an Official Publication of the European Peptide Society·Do-Hyun KimYoon-Sik Lee
Jul 17, 2013·Journal of Thrombosis and Haemostasis : JTH·S Krishnaswamy
Oct 29, 2011·Journal of Cellular Biochemistry·Soumendra RanaAlireza R Rezaie
Jan 12, 2005·Protein Expression and Purification·Charlotte Harkjaer FynboThor Las Holtet
Feb 16, 2006·Neural Networks : the Official Journal of the International Neural Network Society·Zheng Rong YangT Charles Hodgman
Oct 30, 2008·Analytical Biochemistry·Dirk WildeboerRamadan A Abuknesha
Apr 20, 2010·Journal of Molecular Biology·Michael J Page, Enrico Di Cera
Oct 16, 2003·The Journal of Biological Chemistry·Daniel O BeckMichael Kalafatis
Oct 30, 2003·The Journal of Biological Chemistry·Elsa P BianchiniBernard F Le Bonniec
Oct 7, 2005·The Journal of Biological Chemistry·Virginie B Louvain-QuintardBernard F Le Bonniec
Jan 30, 2016·American Journal of Physiology. Gastrointestinal and Liver Physiology·Anita BalázsMiklós Sahin-Tóth
May 10, 2003·The Journal of Biological Chemistry·Lisam S SinghMichael Kalafatis
Apr 26, 2007·The Biochemical Journal·Katrine S LarsenHenning R Stennicke
Dec 23, 2004·The Journal of Physiology·Hye Jin LeeRobert M Corn
Feb 26, 2008·The Journal of Biological Chemistry·Hung-Ju HsuAn-Suei Yang
May 7, 2021·Essays in Biochemistry·Aatir A TungekarLloyd W Ruddock
Jul 30, 2009·Archives of Biochemistry and Biophysics·Natasha May-Yoke NgRobert Neil Pike
Sep 1, 2009·Biochemical and Biophysical Research Communications·Likui YangAlireza R Rezaie

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.