Welcome to Entry Card of Lysosomal Enzymes


Details of RareLED ID Assay_1024
Primary information
GENEGBA
NUMBER OF ASSAY84
ASSAY IDs342801 402241 326301 326300 301611 275320 1259217 1258953 1257697 1189723 1189722 1166038 1152619 1152618 1125775 1125774 1122298 1122297 1122296 1122295 40289 40287 764875 764874 711099 702246 702244 702229 702066 702063 699826 699346 699345 699344 699343 699342 699341 699340 662915 662914 652473 652472 652471 617953 617951 613052 592293 592238 592237 579723 553253 500310 482234 482197 482196 482195 474613 474612 456165 350169 40442 40439 40425 1344544 220899 220895 218556 218548 40444 40424 40423 40288 39995 32963 1322883 1322882 1321659 1321658 1321657 1288864 1288863 1288862 1260570 1268
Assay Name1. Inhibition of human lysosomal beta-glucosidase,2. Inhibition of human placental beta-glucocerebrosidase by spectrophotometry,3. Effect on recombinant wild-type glucocerebrosidase,4. Inhibition of recombinant wild-type glucocerebrosidase,5. Inhibition of glucocerebrosidase,6. Inhibition of glucocerebrosidase,7. Enzyme Assay: To 2.5 A inverted question markl of supernatant (in 96-well plates) was added 17.5 A inverted question markl reaction buffer (citrate phosphate buffer, pH 4.5, no Triton X-100), and 50 A inverted question markl of 4-methyl umbelliferone (4-MU)-labeled substrate, A?-glucopyranoside, or a labeled negative controls (A inverted question mark-glucopyranoside or A inverted question mark-galacatopyranoside). Plates were incubated at 37A degrees for 1 hour, followed by the addition of 70 A inverted question markl stop buffer (0.4 M glycine-NaOH, pH 10.6). Activity of GCase was determined by measuring the emission at 460 nm by exciting at 355 nm using a 1 second read time per well (Victor2 multilabel counter-Wallac) Enzyme activity was normalized to the amount in A inverted question markl of lysate added, and enzyme activity per A inverted question markl of lysate was estimated.,8. Enzyme Activity Assay: Activity of GBA was measured at 37A degrees C. with 4-methylumbelliferyl A?-D-glucopyranoside as substrate as reported previously. To determine the IC50 value, the inhibitors were preincubated for 30 min with the enzyme before addition of the substrate mixture. The incubation mixture contained 3 mM fluorogenic substrate, 0.2% (w/v) sodium taurocholate and 0.1% (v/v) Triton X-100 in 150 mM McIlvaine buffer, pH 5.2. After stopping the incubation with excess NaOH-glycine (pH 10.3), we measured fluorescence with a fluorimeter LS 30 (Perkin Elmer) using A inverted question markex 366 nm and A inverted question markem 445 nm. Activity of lactase was quantified by measuring liberated glucose from lactase38. In vivo activity of GBA in cells was measured using FDG as substrate and FACS32.,9. Enzyme Inhibition Assay: The enzyme inhibition assays used monitored the ability of a test compound to bind and prevent the hydrolysis of a fluorogenic substrate in a concentration-dependent mariner. Specifically, the enzyme activity of recombinant human GCase (rhGCase; Cerezyme, Genzyme Corp.) was measured using the 4-methylumbelliferyl-A?-D-glucopyranoside (4-MU-A?-D-Glc) fluorogenic substrate in the absence or in the presence of varying amounts of each test compound.,10. Inhibition of human beta-glucocerebrosidase at pH 5.5 by spectrophotometry,11. Inhibition of human beta-glucocerebrosidase at pH 7.0 by spectrophotometry,12. Inhibition of GBA1 (unknown origin) using beta-D-[1-14C]glucocerebroside assessed as 4-methylumbelliferrone by fluorimetry,13. Inhibition of human lysosomal beta-glucocerebrosidase using 2,4-dinitrophenyl-beta-D-glucopyranoside as substrate by UV spectrophotometric analysis,14. Inhibition of human lysosomal beta-glucocerebrosidase,15. Competitive inhibition of human lysosomal beta-glucocerebrosidase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 45 mins before substrate addition measured after 30 mins by double-reciprocal plot analysis,16. Inhibition of human lysosomal beta-glucocerebrosidase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 45 mins before substrate addition measured after 30 mins by fluorescence spectrophotometer analysis,17. Inhibition of human recombinant imiglucerase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 30 mins at pH 5.2 before substrate addition measured after 10 mins,18. Inhibition of human recombinant imiglucerase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 30 mins at pH 7 before substrate addition measured after 10 mins,19. Competitive inhibition of human recombinant imiglucerase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 30 mins at pH 5.2 before substrate addition measured after 10 mins by Lineweaver-Burk or Dixon plot analysis,20. Competitive inhibition of human recombinant imiglucerase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 30 mins at pH 7 before substrate addition measured after 10 mins by Lineweaver-Burk or Dixon plot analysis,21. Tested for inhibitory activity against Beta-glucosidase from Agrobacterium species,22. Inhibitory activity against Agrobacterium sp. Beta-glucosidase employing fluorescence spectrometric method,23. Competitive inhibition of human beta-glucocerebrosidase by Lineweaver-Burk double reciprocal plot method,24. Non-competitive inhibition of human beta-glucocerebrosidase by Lineweaver-Burk double reciprocal plot method,25. Inhibition of human beta-glucocerebrosidase,26. Activation of wild type glucocerebrosidase in human spleen homogenate using 4methylumbelliferone-Glc as substrate assessed as half-maximal activation concentration incubated for 5 mins prior to substrate addition measured after 30 to 45 mins by fluorimetric analysis,27. Activation of wild type glucocerebrosidase using 4methylumbelliferone-Glc as substrate assessed as half-maximal activation concentration incubated for 5 mins prior to substrate addition measured after 30 to 45 mins by fluorimetric analysis,28. Binding affinity to NT-647 labeled glucocerebrosidase by microscale thermophoretic analysis,29. Inhibition of wild type glucocerebrosidase using 4-methylumbellifereno-Glc as substrate incubated for 5 mins prior to substrate addition measured after 20 mins by fluorimetric analysis,30. Inhibition of wild type glucocerebrosidase in human spleen homogenate using 4-methylumbellifereno-Glc as substrate incubated for 5 mins prior to substrate addition measured after 20 mins by fluorimetric analysis,31. Inhibition of beta-glucocerebrosidase in human fibroblast lysate using 4-methylumbelliferyl beta-D-glycopyranoside as substrate by fluorimetric analysis,32. Binding affinity to recombinant imiglucerase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 30 mins before substrate addition by Dixon plot assay,33. Binding affinity to recombinant imiglucerase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 30 mins before substrate addition by Lineweaver-Burk plot assay,34. Competitive inhibition of recombinant Imiglucerase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 30 mins before substrate addition at pH 7.0 by Dixon plot assay,35. Competitive inhibition of recombinant Imiglucerase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 30 mins before substrate addition at pH 5.2 by Dixon plot assay,36. Competitive inhibition of recombinant Imiglucerase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 30 mins before substrate addition at pH 7.0 Lineweaver-Burk plot assay,37. Competitive inhibition of recombinant Imiglucerase using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 30 mins before substrate addition at pH 5.2 by Lineweaver-Burk plot assay,38. Inhibition of lysosomal beta-glucosidase in human fibroblast using 4-methylumbelliferyl-beta-D-glucopyranoside as substrate preincubated for 24 hrs before substrate addition measured after 2 hrs by fluorometry,39. Competitive inhibition of wild type human glucocerebrosidase using 4-methylumbelliferyl beta-D-glucopyranoside as substrate after 15 mins by Dixon and Line-Weaver Burk plot analysis at pH 5,40. Competitive inhibition of wild type human glucocerebrosidase using 2,4-dinitrophenyl beta-D-glucopyranoside as substrate after 3 mins by Dixon and Line-Weaver Burk plot analysis at pH 5,41. Competitive inhibition of human recombinant beta-glucocerebrosidase assessed as 4-methyumbelliferone formation after 30 mins by Lineweaver-Burk plot analysis at pH 5.2,42. Inhibition of human recombinant beta-glucocerebrosidase assessed as 4-methyumbelliferone formation after 30 mins by spectrophotometric analysis at pH 5.2,43. Inhibition of human recombinant beta-glucocerebrosidase assessed as 4-methyumbelliferone formation after 30 mins by spectrophotometric analysis at pH 7,44. Inhibition of beta-glucosidase,45. Uncompetitive inhibition of human beta-glucosidase by Michaelis-Menten kinetics analysis,46. Inhibition of human lysosome beta-glucosidase assessed as production of 4-methylumbelliferone using 4-methylumbelliferyl beta-D-glucoside as substrate by spectrophotometry,47. Competitive inhibition of recombinant glucocerebrosidase,48. Inhibition of recombinant glucocerebrosidase in McIlvaine buffer at pH 7.4 after 10 mins by fluorometric analysis,49. Inhibition of recombinant glucocerebrosidase in McIlvaine buffer at pH 5.2 after 10 mins by fluorometric analysis,50. Inhibition of partially purified human placental beta-glucocerebrosidase after 15 to 60 mins,51. Inhibition of recombinant GBA1 preincubated with compound for 30 mins using 4-methylumbelliferyl-B-glucoside substrate by fluorimetric assay,52. Inhibition of GCase assessed as 4-methylumbelliferone release assay after 30 mins by fluorimetry,53. Inhibition of recombinant beta-glucocerebrosidase after 10 mins by fluorimetry,54. Competitive inhibition of beta-glucocerebrosidase at pH 5.2 by Lineweaver-Burke plot analysis,55. Inhibition of recombinant beta-glucocerebrosidase at pH 5.2 after 10 mins by fluorimetry,56. Inhibition of recombinant beta-glucocerebrosidase at pH 7.4 after 10 mins by fluorimetry,57. Inhibition of human recombinant beta-glucocerebrosidase preincubated for 10 mins before 4-methylumbelliferyl-betaD-glucopyranoside addition measured after 10 mins by spectrophotometry,58. Inhibition of human recombinant beta-glucocerebrosidase assessed as p-nitrophenolate accumulation preincubated for 10 mins before p-nitrophenyl-beta-d-glucopyranoside addition measured after 10 mins by fluoresce
ACTIVITYSubstance BioActivity: 3 Active, 1 Activity <= 1 µM, 5 Tested Substance BioActivity: 1 Tested Substance BioActivity: 7 Active, 4 Activity <= 1 µM, 25 Tested Substance BioActivity: 21 Active, 13 Activity <= 1 µM, 27 Tested Substance BioActivity: 36 Active, 6 Activity <= 1 µM, 50 Tested Substance BioActivity: 2 Active, 5 Tested Substance BioActivity: 9 Active, 20 Tested Substance BioActivity: 4 Active, 3 Activity <= 1 µM, 4 Tested Substance BioActivity: 7 Active, 7 Activity <= 1 µM, 7 Tested Substance BioActivity: 1 Active, 6 Tested Substance BioActivity: 2 Active, 6 Tested Substance BioActivity: 67 Active, 35 Activity <= 1 µM, 77 Tested Substance BioActivity: 2 Active, 1 Activity <= 1 µM, 3 Tested Substance BioActivity: 3 Active, 1 Activity <= 1 nM, 2 Activity <= 1 µM, 4 Tested Substance BioActivity: 2 Active, 4 Tested Substance BioActivity: 2 Active, 4 Tested Substance BioActivity: 3 Active, 1 Activity <= 1 µM, 4 Tested Substance BioActivity: 3 Active, 3 Activity <= 1 µM, 4