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The human ENO1 gene product (recombinant human ��-enolase) displays characteristics required for a plasminogen binding protein

Journal Article


Abstract


  • Plasminogen binds with low affinity in a lysine-dependent manner to many cell types. Previously, a 54 kDa plasminogen receptor found on the surface of U-937 cells was identified as an ��-enolase-like molecule. The aims of this study were to determine whether recombinant ��-enolase (r-��-enolase), encoded by ENO1, was a plasminogen binding protein and to generate polyclonal antibodies against this antigen. Plasminogen specifically bound r-��-enolase with a K(d) 1.9 ��M and approached saturation at 10 ��M. Lysine-dependent plasminogen binding to r-��-enolase was demonstrated by a greater than 80% inhibition of binding by the lysine analogues ��-amino caproic acid and tranexamic acid, whilst only 14% inhibition occurred with the arginine analogue benzamidine. Removal of the C-terminal lysine residue of r-��-enolase with carboxypeptidase B significantly reduced its plasminogen binding capacity, suggesting that binding required C-terminal lysine residue of r-��-enolase. Binding to r-��-enolase enhanced the activation rate of plasminogen by urokinase but prevented ��2-antiplasmin from binding plasminogen. Taken together, these data suggest that the gene product of human ENO1 encodes an authentic plasminogen binding protein.

Publication Date


  • 1997

Citation


  • Andronicos, N. M., Ranson, M., Bognacki, J., & Baker, M. S. (1997). The human ENO1 gene product (recombinant human ��-enolase) displays characteristics required for a plasminogen binding protein. Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology, 1337(1), 27-39. doi:10.1016/S0167-4838(96)00146-X

Scopus Eid


  • 2-s2.0-0031552061

Start Page


  • 27

End Page


  • 39

Volume


  • 1337

Issue


  • 1

Place Of Publication


Abstract


  • Plasminogen binds with low affinity in a lysine-dependent manner to many cell types. Previously, a 54 kDa plasminogen receptor found on the surface of U-937 cells was identified as an ��-enolase-like molecule. The aims of this study were to determine whether recombinant ��-enolase (r-��-enolase), encoded by ENO1, was a plasminogen binding protein and to generate polyclonal antibodies against this antigen. Plasminogen specifically bound r-��-enolase with a K(d) 1.9 ��M and approached saturation at 10 ��M. Lysine-dependent plasminogen binding to r-��-enolase was demonstrated by a greater than 80% inhibition of binding by the lysine analogues ��-amino caproic acid and tranexamic acid, whilst only 14% inhibition occurred with the arginine analogue benzamidine. Removal of the C-terminal lysine residue of r-��-enolase with carboxypeptidase B significantly reduced its plasminogen binding capacity, suggesting that binding required C-terminal lysine residue of r-��-enolase. Binding to r-��-enolase enhanced the activation rate of plasminogen by urokinase but prevented ��2-antiplasmin from binding plasminogen. Taken together, these data suggest that the gene product of human ENO1 encodes an authentic plasminogen binding protein.

Publication Date


  • 1997

Citation


  • Andronicos, N. M., Ranson, M., Bognacki, J., & Baker, M. S. (1997). The human ENO1 gene product (recombinant human ��-enolase) displays characteristics required for a plasminogen binding protein. Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology, 1337(1), 27-39. doi:10.1016/S0167-4838(96)00146-X

Scopus Eid


  • 2-s2.0-0031552061

Start Page


  • 27

End Page


  • 39

Volume


  • 1337

Issue


  • 1

Place Of Publication