β-Galactosidase treatment is a common first-stage modification of the three major subtypes of Gc protein to GcMAF.
BACKGROUND
The 1f1f subtype of the group-specific component (Gc) protein is converted into Gc protein-derived macrophage-activating factor (GcMAF) by enzymatic processing with β-galactosidase and sialidase. We previously demonstrated that preGc(1f1f)MAF, a full Gc(1f1f) protein otherwise lacking a galactosyl moiety, can be converted to GcMAF by treatment with mouse peritoneal fluid. Here, we investigated the effects of the β-galactosidase-treated 1s1s and 22 subtypes of Gc protein (preGc(1s1s)MAF and preGc₂₂MAF) on the phagocytic activation of mouse peritoneal macrophages.


RESULTS
We demonstrated the presence of Gal-GalNAc disaccharide sugar structures in the Gc(1s1s) protein by western blotting using peanut agglutinin and Helix pomatia agglutinin lectin. We also found that preGc(1s1s)MAF and preGc₂₂MAF significantly enhanced the phagocytic activity of mouse peritoneal macrophages in the presence and absence of mouse peritoneal fluid.


CONCLUSIONS
We demonstrate that preGc(1s1s)MAF and preGc₂₂MAF proteins can be used as effective macrophage activators.
Department of Life System, Institute of Technology and Science, Graduate School, The University of Tokushima, 2-1 Minamijosanjimacho, Tokushima 770-8506, Japan. uto@bio.tokushima-u.ac.jp
Yoshihiro
Y
Uto
Syota
S
Yamamoto
Hirotaka
H
Mukai
Noriko
N
Ishiyama
Ryota
R
Takeuchi
Yoshinori
Y
Nakagawa
Keiji
K
Hirota
Hiroshi
H
Terada
Shinya
S
Onizuka
Hitoshi
H
Hori
D000818:Animals; D015153:Blotting, Western; D006801:Humans; D008262:Macrophage Activation / Q000502:physiology*; D016215:Macrophage-Activating Factors / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D010587:Phagocytosis / Q000502:physiology; D014809:Vitamin D-Binding Protein / Q000378:metabolism*; D001616:beta-Galactosidase / Q000378:metabolism*
D016215:Macrophage-Activating Factors; D014809:Vitamin D-Binding Protein; D001616:beta-Galactosidase