Use of beta-maltosides (p-nitrophenyl-beta-D-maltoside, 2-chloro-4- nitrophenyl-beta-D-maltoside and 4-methylumbelliferyl-beta-D-maltoside) as substrates for the assay of neutral alpha-glucosidase from human kidney and urine.
The method of assay of neutral alpha-glucosidase from human kidney and urine using beta-maltosides (p-nitrophenyl-beta-D-maltoside [NP-beta-D-maltoside], 2-chloro-4-nitrophenyl-beta-D-maltoside]) [CNP-beta-D-maltoside] and 4-methylumbelliferyl-beta-D-maltosides ([MU-beta-D-maltoside]) as substrates and beta-glucosidase as an auxiliary enzyme is proposed. All three beta-maltosides are suitable substrates for the determination of neutral alpha-glucosidase activity but MU-beta-D-maltoside is the most sensitive due to its methylumbelliferyl moiety. The method is simple, convenient and 10-fold more sensitive than the commonly used alpha-glucosidase assay procedure with the corresponding synthetic alpha-glucosides, p-nitrophenyl- alpha-D-glucoside (NP-alpha-D-glucoside) and 4-methylumbelliferyl-alpha-D-glucoside (MU-alpha-D-glucoside). A modification of the method, with p-nitrophenyl-beta-D-maltoside as substrate, was applied to the semiautomatic assay of urinary alpha-glucosidase in 96-well microtitre plates.
Lukomskaya
I S
IS
Institute of Biomedical Chemistry of RAMS, Moscow, Russian Federation.
Voznyi
Y V
YV
Lanskaya
I M
IM
Podkidisheva
E I
EI
D000118:Acetylglucosaminidase / Q000652:urine; D001331:Automation; D005960:Glucosides; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives; D007668:Kidney / Q000201:enzymology*; D016030:Kidney Transplantation / Q000502:physiology*; D007700:Kinetics; D008320:Maltose / Q000031:analogs & derivatives; D008871:Microvilli / Q000201:enzymology; D009578:Nitrobenzenes; D012680:Sensitivity and Specificity; D013053:Spectrophotometry / Q000379:methods; D013379:Substrate Specificity; D000519:alpha-Galactosidase / Q000032:analysis / Q000378:metabolism* / Q000652:urine
C099582:4-methylumbelliferylmaltoside; D005960:Glucosides; D009578:Nitrobenzenes; C078908:2-chloro-4-nitrophenylmaltoside; C036988:4-nitrophenylmaltoside; D006923:Hymecromone; D008320:Maltose; D000519:alpha-Galactosidase; D000118:Acetylglucosaminidase