Figure 9.
Replacement of RP S19-like molecule in plasma with recombinant RP S19 and with mutants of RP S19. PPP was pretreated with the anti-RP S19 rabbit IgG beads as Figure 4. Aliquots of it were reconstituted with a recombinant wild-type RP S19 (RPS19-re/) at concentrations of 1 or of 10 μg/ml, or with one of the RP S19 mutants, Gln137Asn-RP S19 (M12-re/), or Lys23, 24, 27, 29Ala-RP S19 (M13-re/) at a concentration of 10 μg/ml and were further added with platelets to reconstitute the PRP. After being coagulated with CA-1, the sera derived from the reconstituted PRP aliquots were heat treated (PRP-S) and subjected to the monocyte chemotaxis assay at serial log-scale dilutions. Positive control for the experiment was a serum derived from normal PRP compensated for the volume change with PBS (PBS/PRP-S). Positive and negative controls for the chemotaxis assay were 10−9 M C5a and PBS, respectively.