Further characterization of the binding of plasminogen to heparin: evidence for the involvement of lysine residues.
We have previously demonstrated that the heparin-binding site of plasminogen is located in Val442-plasminogen region (kringle 5 domain plus light (B) chain) (Soeda, S., Kakiki, M., Shimeno, H., and Nagamatsu, A. (1987) Biochim. Biophys. Acta 916, 279-287). The chemical modification of Val442-plasminogen with a lysine reagent, pyridoxal 5'-phosphate (PLP), and sodium borohydride resulted in the incorporation of 8-10 PLP moieties per molecule of the zymogen. This PLP-labeled zymogen had no affinity for a heparin-Sepharose column, whereas the non-labeled one bound to the column. Modification in the presence of heparin decreased the extent of labeling by 1-2 mol of PLP per mol of Val442-plasminogen. To further examine the binding site of plasminogen to heparin, functionally active A and B chains were separated from Lys-plasmin after mild reduction and S-carboxymethylation. Only B chain possessed affinity for heparin-Sepharose. Furthermore, plasmin(ogen) bound to heparin was protected from alpha 2-antiplasmin inhibition. These results indicate that one or two lysine residues located in the catalytic region (B chain) of plasmin(ogen) are essential to heparin binding, and that the binding of plasminogen to heparin or heparin-like substance in extracellular matrix environments may be important for the localization and activation of plasminogen and for the prolongation of the resultant plasmin activity.
Soeda
S
S
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Fukuoka University, Japan.
Ohki
H
H
Shimeno
H
H
Nagamatsu
A
A
D001665:Binding Sites; D001894:Borohydrides; D055598:Chemical Phenomena; D002621:Chemistry; D005341:Fibrinolysin / Q000378:metabolism; D005971:Glutamates / Q000378:metabolism; D018698:Glutamic Acid; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D008239:Lysine; D010958:Plasminogen / Q000378:metabolism*; D011732:Pyridoxal Phosphate; D014633:Valine
D001894:Borohydrides; D005971:Glutamates; D018698:Glutamic Acid; D011732:Pyridoxal Phosphate; C025364:sodium borohydride; D010958:Plasminogen; D006493:Heparin; D005341:Fibrinolysin; D014633:Valine; D008239:Lysine