Neutral alpha-glucosidase in granule fractions from guinea pig polymorphonuclear leukocytes: enzymic characterization and comparative studies with monoclonal antibodies.
Neutral alpha-glucosidase was partially purified from granular fractions isolated from guinea pig polymorphonuclear leukocytes (PMNL). The native enzyme had a high molecular weight, about 417,000, with a subunit of 43,000. The purified enzyme hydrolysed 4-methylumbelliferyl alpha-glucoside and maltose, but not isomaltose, trehalose, and glycogen. The enzyme was strongly inhibited by bromoconduritol and castanospermine, but only slightly by turanose. Monoclonal antibodies which can bind specifically to the enzyme were prepared by immunizing mice with the partially purified enzyme. Hybridomas producing the monoclonal antibodies were selected by an enzyme-linked immunosorbent assay. The seven monoclonal antibodies were found to react with the enzyme from PMNL, but not with the glycoprotein-processing alpha-glucosidase isolated from liver microsomes nor with the macrophage enzyme. The results indicated that PMNL contain a particulate neutral alpha-glucosidase enzymologically and immunologically distinct from other alpha-glucosidases.
Imai
K
K
Department of Biochemistry, Shimane Medical University.
Harada
T
T
Takano
Y
Y
Morikawa
S
S
Tanaka
A
A
D000818:Animals; D000911:Antibodies, Monoclonal; D002850:Chromatography, Gel; D003594:Cytoplasmic Granules / Q000201:enzymology; D004797:Enzyme-Linked Immunosorbent Assay; D006168:Guinea Pigs; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008862:Microsomes, Liver / Q000201:enzymology; D008970:Molecular Weight; D009504:Neutrophils / Q000201:enzymology* / Q000276:immunology; D011489:Protein Denaturation; D000520:alpha-Glucosidases / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000520:alpha-Glucosidases