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Structural basis of Na+ activation mimicry in murine thrombin.

Marino F, Chen ZW, Ergenekan CE, Bush-Pelc LA, Mathews FS, Di Cera E
J Biol Chem 282 16355-61 (2007)
Cited: 8 times
EuropePMC logo PMID: 17428793

Abstract

Unlike human thrombin, murine thrombin lacks Na+ activation due to the charge reversal substitution D222K in the Na+ binding loop. However, the enzyme is functionally stabilized in a Na+-bound form and is highly active toward physiologic substrates. The structural basis of this peculiar property is unknown. Here, we present the 2.2 A resolution x-ray crystal structure of murine thrombin in the absence of inhibitors and salts. The enzyme assumes an active conformation, with Ser-195, Glu-192, and Asp-189 oriented as in the Na+-bound fast form of human thrombin. Lys-222 completely occludes the pore of entry to the Na+ binding site and positions its side chain inside the pore, with the Nzeta atom H-bonded to the backbone oxygen atoms of Lys-185, Asp-186b, and Lys-186d. The same architecture is observed in the 1.75 A resolution structure of a thrombin chimera in which the human enzyme carries all residues defining the Na+ pore in the murine enzyme. These findings demonstrate that Na+ activation in thrombin is linked to the architecture of the Na+ pore. The molecular strategy of Na+ activation mimicry unraveled for murine thrombin is relevant to serine proteases and enzymes activated by monovalent cations in general.

Reviews citing this publication (3)

  1. Thrombin. Di Cera E. Mol Aspects Med 29 203-254 (2008)
  2. Conformational selection in trypsin-like proteases. Pozzi N, Vogt AD, Gohara DW, Di Cera E. Curr Opin Struct Biol 22 421-431 (2012)
  3. Allostery in trypsin-like proteases suggests new therapeutic strategies. Gohara DW, Di Cera E. Trends Biotechnol 29 577-585 (2011)

Articles citing this publication (5)

  1. Structural identification of the pathway of long-range communication in an allosteric enzyme. Gandhi PS, Chen Z, Mathews FS, Di Cera E. Proc Natl Acad Sci U S A 105 1832-1837 (2008)
  2. Crystal structures of murine thrombin in complex with the extracellular fragments of murine protease-activated receptors PAR3 and PAR4. Bah A, Chen Z, Bush-Pelc LA, Mathews FS, Di Cera E. Proc Natl Acad Sci U S A 104 11603-11608 (2007)
  3. Allosterism-based simultaneous, dual anticoagulant and antiplatelet action: allosteric inhibitor targeting the glycoprotein IbĪ±-binding and heparin-binding site of thrombin. Mehta AY, Mohammed BM, Martin EJ, Brophy DF, Gailani D, Desai UR. J Thromb Haemost 14 828-838 (2016)
  4. Mechanism of the anticoagulant activity of thrombin mutant W215A/E217A. Gandhi PS, Page MJ, Chen Z, Bush-Pelc L, Di Cera E. J Biol Chem 284 24098-24105 (2009)
  5. Rigidification of the autolysis loop enhances Na(+) binding to thrombin. Pozzi N, Chen R, Chen Z, Bah A, Di Cera E. Biophys Chem 159 6-13 (2011)


Related citations provided by authors (1)

  1. MOLECULAR DISSECTION OF Na+ BINDING TO THROMBIN.. Pineda AO, Carrell CJ, Bush LA, Prasad S, Caccia S, Chen ZW, Mathews FS, Di Cera E J. Biol. Chem. 279 31842- (2004)

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