Glycoprotein receptors to wheat germ agglutinin on the surface of murine macrophages.
Peritoneal macrophages elicited in various ways exhibited marked wheat germ agglutinin (WGA)-dependent cytolytic activity against MM46 cells [nearly 100% lysis at effector to target ratio (E/T) = 40], while macrophage cell lines J774.1 and WEHI-3 cells showed less activity (-25% at E/T = 0.63-2.5) and no cytolysis (E/T = 40), respectively. Studies on the kinetics of WGA-binding showed that peritoneal macrophages had an association constant (Ka) of 0.09-0.18 X 10(6) M-1 and the number of their binding sites (N) was 2-7.7 X 10(7)/cell, while the Ka and N values of J774.1 and WEHI-3 cells were 1.2 X 10(6) M-1 and 4.1 X 10(7)/cell and 0.6 X 10(6) M-1 and 5.4 X 10(7)/cell, respectively. On the other hand, cytolytic target MM46 cells showed values of 0.27 X 10(6) M-1 and 1.8 X 10(8)/cell. Lower affinities and moderate numbers of binding sites coincided with the efficient WGA-dependent cytolytic activities of the effector cells. The surface of peritoneal macrophages and J774.1 and MM46 cells was iodinated, the cells lysed, the lysates fractionated on a WGA-Sepharose column and the WGA-binding surface-glycoproteins in GlcNAc-specific fractions of the eluate were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Peritoneal macrophages gave three major bands of 153 kilo dalton (kDa), 140 and 90 kDa proteins, whereas J774.1 and MM46 cells both gave 6 bands of very similar molecular weights and intensities that were quite unlike those of peritoneal macrophages.(ABSTRACT TRUNCATED AT 250 WORDS)
Satoh
M
M
Faculty of Pharmaceutical Sciences, Teikyo University, Kanagawa, Japan.
Yamazaki
M
M
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D005122:Exudates and Transudates / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D066298:In Vitro Techniques; D008264:Macrophages / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D011975:Receptors, Mitogen / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000378:metabolism
D006023:Glycoproteins; D011975:Receptors, Mitogen; C027503:wheat germ agglutinin receptor