The chemokine macrophage inflammatory protein (MIP)-2 alpha was identified
as a plasminogen binding protein by phage display analysis. MIP-2 alpha and
a truncated form lacking 5 lysine residues in the COOH-terminal region (mu
t-MIP-2 alpha) were expressed in E. coli and purified to apparent homogenei
ty, Purified MIP-2 alpha but not mut-MIP-2 alpha. bound specifically to pla
sminogen. with K-A of 3.7 x 10(9) M-1 for the interaction of plasminogen wi
th surface-bound MIP-2 alpha. Binding and competition experiments indicated
that the interaction involves the region comprising the first 3 kringles o
f plasminogen and the COOH-terminal lysine-rich domain of MIP-2 alpha. Acti
vation of plasminogen bound to surface-associated MIP-2 alpha by two-chain
urokinase-type plasminogen activator (tcu-PA) was about 2.5-fold more effic
ient than in solution (catalytic efficiency k(cat)/K-M of 0.1 mu M(-1)s(-1)
as compared to 0.04 mu M(-1)s(-1)). In contrast, binding of plasminogen to
MIP-2 alpha in solution was very weak. as evidenced by the absence of comp
etition of MIP-2 alpha with lysine-Sepharose or with human THP-1 cells for
binding of plasminogen. In agreement with this finding, addition of excess
MIP-2 alpha did not affect the main functional properties of plasmin(ogen)
in solution, as indicated by unaltered activation rates of plasminogen by t
cu-PA or tissue-type plasminogen activator (t-PA), t-PA-mediated fibrinolys
is, and inhibition rate of plasmin by alpha(2)-antiplasmin. Thus, associati
on of MIP-2 alpha with surfaces exposes its COOH-terminal plasminogen-bindi
ng site, and may result in enhanced local plasmin generation.