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Autoactivation of thrombin precursors.

Pozzi N, Chen Z, Zapata F, Niu W, Barranco-Medina S, Pelc LA, Di Cera E
J Biol Chem 288 11601-10 (2013)
Related entries: 3sqe, 3sqh, 4h6s, 4h6t, 4hfp

Cited: 30 times
EuropePMC logo PMID: 23467412

Abstract

Trypsin-like proteases are synthesized as inactive zymogens and convert to the mature form upon activation by specific enzymes, often assisted by cofactors. Central to this paradigm is that the zymogen does not convert spontaneously to the mature enzyme, which in turn does not feed back to activate its zymogen form. In the blood, the zymogens prothrombin and prethrombin-2 require the prothrombinase complex to be converted to the mature protease thrombin, which is unable to activate prothrombin or prethrombin-2. Here, we show that replacement of key residues within the activation domain causes these zymogens to spontaneously convert to thrombin. The conversion is started by the zymogen itself, which is capable of binding ligands at the active site, and is abrogated by inactivation of the catalytic residue Ser-195. The product of autoactivation is functionally and structurally equivalent to wild-type thrombin. Zymogen autoactivation is explained by conformational selection, a basic property of the trypsin fold uncovered by structural and rapid kinetics studies. Both the zymogen and protease undergo a pre-existing equilibrium between active and inactive forms. The equilibrium regulates catalytic activity in the protease and has the potential to unleash activity in the zymogen to produce autoactivation. A new strategy emerges for the facile production of enzymes through zymogen autoactivation that is broadly applicable to trypsin-like proteases of biotechnological and clinical interest.

Reviews - 4rn6 mentioned but not cited (1)

  1. Exosite Binding in Thrombin: A Global Structural/Dynamic Overview of Complexes with Aptamers and Other Ligands. Troisi R, Balasco N, Autiero I, Vitagliano L, Sica F. Int J Mol Sci 22 10803 (2021)


Reviews citing this publication (2)

  1. Proteolytic activity of contact factor zymogens. Shamanaev A, Emsley J, Gailani D. J Thromb Haemost 19 330-341 (2021)
  2. Structure of Coagulation Factor II: Molecular Mechanism of Thrombin Generation and Development of Next-Generation Anticoagulants. Chinnaraj M, Planer W, Pozzi N. Front Med (Lausanne) 5 281 (2018)

Articles citing this publication (27)

  1. Histone H4 promotes prothrombin autoactivation. Barranco-Medina S, Pozzi N, Vogt AD, Di Cera E. J Biol Chem 288 35749-35757 (2013)
  2. Crystal structure of prothrombin reveals conformational flexibility and mechanism of activation. Pozzi N, Chen Z, Gohara DW, Niu W, Heyduk T, Di Cera E. J Biol Chem 288 22734-22744 (2013)
  3. The linker connecting the two kringles plays a key role in prothrombin activation. Pozzi N, Chen Z, Pelc LA, Shropshire DB, Di Cera E. Proc Natl Acad Sci U S A 111 7630-7635 (2014)
  4. Kinetic dissection of the pre-existing conformational equilibrium in the trypsin fold. Vogt AD, Chakraborty P, Di Cera E. J Biol Chem 290 22435-22445 (2015)
  5. How the Linker Connecting the Two Kringles Influences Activation and Conformational Plasticity of Prothrombin. Pozzi N, Chen Z, Di Cera E. J Biol Chem 291 6071-6082 (2016)
  6. Structure of prothrombin in the closed form reveals new details on the mechanism of activation. Chinnaraj M, Chen Z, Pelc LA, Grese Z, Bystranowska D, Di Cera E, Pozzi N. Sci Rep 8 2945 (2018)
  7. Structural Architecture of Prothrombin in Solution Revealed by Single Molecule Spectroscopy. Pozzi N, Bystranowska D, Zuo X, Di Cera E. J Biol Chem 291 18107-18116 (2016)
  8. An allosteric switch for pro-HGF/Met signaling using zymogen activator peptides. Landgraf KE, Steffek M, Quan C, Tom J, Yu C, Santell L, Maun HR, Eigenbrot C, Lazarus RA. Nat Chem Biol 10 567-573 (2014)
  9. Dynamics Govern Specificity of a Protein-Protein Interface: Substrate Recognition by Thrombin. Fuchs JE, Huber RG, Waldner BJ, Kahler U, von Grafenstein S, Kramer C, Liedl KR. PLoS One 10 e0140713 (2015)
  10. Interplay between conformational selection and zymogen activation. Chakraborty P, Acquasaliente L, Pelc LA, Di Cera E. Sci Rep 8 4080 (2018)
  11. Role of the I16-D194 ionic interaction in the trypsin fold. Stojanovski BM, Chen Z, Koester SK, Pelc LA, Di Cera E. Sci Rep 9 18035 (2019)
  12. Residues W215, E217 and E192 control the allosteric E*-E equilibrium of thrombin. Pelc LA, Koester SK, Chen Z, Gistover NE, Di Cera E. Sci Rep 9 12304 (2019)
  13. Probing prothrombin structure by limited proteolysis. Acquasaliente L, Pelc LA, Di Cera E. Sci Rep 9 6125 (2019)
  14. WEDGE: an anticoagulant thrombin mutant produced by autoactivation. Wood DC, Pelc LA, Pozzi N, Wallisch M, Verbout NG, Tucker EI, Gruber A, Di Cera E. J Thromb Haemost 13 111-114 (2015)
  15. Structural transitions during prothrombin activation: On the importance of fragment 2. Adams TE, Huntington JA. Biochimie 122 235-242 (2016)
  16. Blocking the proteolytic activity of zymogen matriptase with antibody-based inhibitors. Tamberg T, Hong Z, De Schepper D, Skovbjerg S, Dupont DM, Vitved L, Schar CR, Skjoedt K, Vogel LK, Jensen JK. J Biol Chem 294 314-326 (2019)
  17. Role of the activation peptide in the mechanism of protein C activation. Stojanovski BM, Pelc LA, Di Cera E. Sci Rep 10 11079 (2020)
  18. Dual effect of histone H4 on prothrombin activation. Pozzi N, Di Cera E. J Thromb Haemost 14 1814-1818 (2016)
  19. The active site region plays a critical role in Na+ binding to thrombin. Pelc LA, Koester SK, Kukla CR, Chen Z, Di Cera E. J Biol Chem 298 101458 (2022)
  20. Thrombin Exosite Maturation and Ligand Binding at ABE II Help Stabilize PAR-Binding Competent Conformation at ABE I. Billur R, Sabo TM, Maurer MC. Biochemistry 58 1048-1060 (2019)
  21. A Novel ELISA Assay for the Detection of Anti-Prothrombin Antibodies in Antiphospholipid Syndrome Patients at High Risk of Thrombosis. Chinnaraj M, Pengo V, Pozzi N. Front Immunol 12 741589 (2021)
  22. Letter Do the crystallographic forms of prethrombin-2 revert to a single form in solution? Wu S, Shim JY, Lee CJ, Pedersen LG. Biophys Chem 203-204 28-32 (2015)
  23. Loop-driven conformational transition between the alternative and collapsed form of prethrombin-2: targeted molecular dynamics study. Wu S. J Biomol Struct Dyn 35 119-127 (2017)
  24. Rational Design of Protein C Activators. Barranco-Medina S, Murphy M, Pelc L, Chen Z, Di Cera E, Pozzi N. Sci Rep 7 44596 (2017)
  25. Cloning, Expression and Purification of Full-length Recombinant Ecarin and Comparing Its Expression and Function with Its Truncated Form. Jafari Z, Bandehpour M, Gheflat S, Mohammadi N, Kazemi B. Iran J Pharm Res 21 e123791 (2022)
  26. New insight into the traditional model of the coagulation cascade and its regulation: illustrated review of a three-dimensional view. Troisi R, Balasco N, Autiero I, Sica F, Vitagliano L. Res Pract Thromb Haemost 7 102160 (2023)
  27. Roles of the clip domains of two protease zymogens in the coagulation cascade in horseshoe crabs. Yamashita K, Shibata T, Takahashi T, Kobayashi Y, Kawabata SI. J Biol Chem 295 8857-8866 (2020)


Related citations provided by authors (1)

  1. Crystal structures of prethrombin-2 reveal alternative conformations under identical solution conditions and the mechanism of zymogen activation.. Pozzi N, Chen Z, Zapata F, Pelc LA, Barranco-Medina S, Di Cera E Biochemistry 50 10195-202 (2011)

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