Glycosylphosphatidylinositol-anchored NAD glycohydrolase is released from peritoneal macrophages activated by interferon-gamma and lipopolysaccharide.
We have previously shown that an ectoenzyme, NAD glycohydrolase (NADase) could be solubilized by treatment with bacterial phosphatidylinositol phospholipase C (PIPLC). However, it is unknown whether endogenous PIPLC can cleave this ectoenzyme. In this study, we used mouse peritoneal exudate macrophages which have been known to have relatively high activity of NADase. The results show that release of ecto-NADase was markedly increased when mouse peritoneal macrophages were costimulated with interferon-gamma (IFN-gamma) and bacterial lipopolysaccharide (LPS), compared to unstimulated cells. This increase was preceded by markedly enhanced activity of endogenous glycosylphosphatidylinositol phospholipase C (GPIPLC). The cross-reacting determinant (CRD) of the glycosylphosphatidylinositol anchor in released NADase from activated macrophages was detected by immunoblotting with anti-CRD antibody. Taken together, ecto-NADase is release from peritoneal exudate macrophages during IFN-gamma/LPS-induced activation and endogenous GPIPLC is involved in the NADase release from the activated macrophages.
Han
M K
MK
Department of Biochemistry, Chonbuk National University Medical School, Chonju, Korea.
Yim
C Y
CY
An
N H
NH
Kim
H R
HR
Kim
U H
UH
D000818:Animals; D004789:Enzyme Activation; D005122:Exudates and Transudates / Q000166:cytology; D005260:Female; D017261:Glycosylphosphatidylinositols / Q000032:analysis / Q000378:metabolism*; D015151:Immunoblotting; D007371:Interferon-gamma / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008262:Macrophage Activation / Q000187:drug effects* / Q000502:physiology*; D017737:Macrophages, Peritoneal / Q000187:drug effects* / Q000201:enzymology* / Q000378:metabolism; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D009244:NAD+ Nucleosidase / Q000378:metabolism*; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism
D017261:Glycosylphosphatidylinositols; D008070:Lipopolysaccharides; D007371:Interferon-gamma; D010727:Phosphoric Diester Hydrolases; D009244:NAD+ Nucleosidase; D043265:Phosphatidylinositol Diacylglycerol-Lyase