Abstract
Using type IV collagen-Sepharose affinity chromatography, the binding proteins were isolated from 125I surface-labeled human neutrophils. SDS-polyacrylamide gel electrophoresis analysis showed that the binding molecules were composed of the 28-, 49-, 67-, and 95-kDa proteins. Western blot analysis revealed that the 95-kDa proteins contained both L-selectin and nonspecific cross-reacting antigen 90 (NCA90), and that the 67-kDa protein was corresponded to 67-kDa elastin/laminin receptor (67BP) . When f-Met-Leu-Phe (fMLP) -stimulated neutrophils were analyzed using the affinity chromatography, the amounts of the binding proteins were increased about three-fold. However, western blot analysis demonstrated that L-selectin was significantly decreased by the fMLP stimulation. On the other hand, NCA90 and 67BP were increased by the stimulation. Together these observations indicate that neutrophils have several kinds of the adhesion molecules to type IV collagen, and that L-selectin is likely important for the binding of resting neutrophils to type IV collagen, whereas 67BP and NCA90 are likely important for the binding of stimulated neutrophils to type IV collagen.
