Mechanism of neutrophil chemiluminescence induced by wheat germ agglutinin:
partial characterization of the antigens recognized by wheat germ
agglutinin
Y Ozaki, J Iwata and T Ohashi
Wheat germ agglutinin (WGA) stimulated neutrophils to produce significant
levels of luminol-dependent chemiluminescence (CL). Since WGA is known to
bind N-acetylglucosamine (GlcNAc) oligomers and N- acetylneuraminic acid
(NANA), we attempted to determine which binding property of WGA is
essential for induction of CL. The succinylated form of WGA (SuWGA), which
is no longer able to bind NANA, was still able to induce CL.
N-Acetylglucosamine at a concentration of 20 mmol/L almost completely
inhibited WGA-induced CL production by neutrophils, whereas bovine
submaxillary gland mucin, a potent blocker of NANA binding of WGA, failed
to inhibit CL production. Lectins with the GlcNAc-binding property were
examined for their ability to induce CL. Those that have higher valences
and have a tendency to bind GlcNAc oligomers in the internal portion of
glycoconjugates were able to induce CL, whereas those that have low
valences and bind terminal GlcNAc of glycoconjugates failed to induce CL
even at high concentrations. Attempts were made to characterize the
neutrophil membrane proteins recognized by WGA. Glycoproteins with a
molecular weight of 25,000 daltons were identified by a 50 mmol/L GlcNAc
elution of WGA gels loaded with 125I-labeled neutrophil membrane proteins.
Elution with 500 mumol/L GlcNAc trimer produced several glycoproteins of
different molecular weights in addition to the glycoproteins of 25,000
daltons. 125I-labeled WGA and SuWGA were used for autoradiographic analysis
of cell extracts of the neutrophils separated on sodium dodecyl sulfate
polyacrylamide gels. WGA recognized multiple glycoproteins of different
molecular weights, whereas SuWGA bound only a few of them. Glycoproteins of
25,000 daltons, probably corresponding to those identified by 50 mmol/L
GlcNAc elution, were also recognized.
Volume 64,
Issue 5,
pp. 1094-1102,
11/01/1984
Copyright © 1984 by The American Society of Hematology
