{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T08:19:08Z","timestamp":1789028348840,"version":"build-2803163510"},"reference-count":0,"publisher":"S. Karger AG","license":[{"start":{"date-parts":[[2017,8,11]],"date-time":"2017-08-11T00:00:00Z","timestamp":1502409600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/karger.com\/pages\/terms-and-conditions"},{"start":{"date-parts":[[2017,8,11]],"date-time":"2017-08-11T00:00:00Z","timestamp":1502409600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/karger.com\/pages\/terms-and-conditions"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Enzyme"],"abstract":"<jats:p>Comparative kinetic studies with glycon inhibitors were used to investigate the\nproperties of the active site of human acid \u00df-glucosidase (EC 3.2.1.45) from normal placenta\nand spleens of type 1 Ashkenazi Jewish Gaucher disease (AJGD) patients. With the pure normal\nenzyme, the specificity of glycon binding was assessed with 35 glucose derivatives and epimers.\nMost glycons were mixed type inhibitors with a predominantly competitive nature (i.e., K(is) &lt; &lt;\nK(ii)) and had low apparent affinity for the enzyme (K(is)app = 20-500 mmol\/l). \u00df-Glucose-\n1-phosphate was unusual, since it inhibited 4-methylumbelliferyl-\u00df-glucoside hydrolysis in an\nuncompetitive pattern (K(i)app = 0.55 mmol\/l) but had no effect on glucosyl ceramide hydrolysis.\nC-1- (1-deoxy-1-amino-\u00df-D-glucose) and C-3- (3-deoxy-3-amino-D-glucose) amino and C-5-\nimino [1-deoxynojirimycin (dNM), nojirimycin and castanospermine] substituted sugars were\nhighly potent inhibitors with K(is)app(\u00df-glucose)\/K(is)app \u2245 10^3-10^5; an amine at C-2 did not alter\nK(is)app compared to \u00df-glucose. The variation of K(is)app with pH for the 5-imino- and 1-\ndeoxy-1-aminoglycosides conformed to a model for the unprotonated inhibitors binding to the\nprotonated forms (EH and EH(2)) of the diprotic (V(max)app and V(max)app\/K(m)app) normal enzyme\n(pK(1) = 4.7; pK(2) = 6.7) with pH-independent K(is)app values of 2.9-9.0 \u00b5mol\/l and 0.22 mmol\/l,\nrespectively. Several of the amine-containing inhibitors competitively protected the enzyme\nfrom inactivation by conduritol B epoxide, a covalent active site-directed inhibitor, indicating\ninteraction with residues at that site. With the partially purified AJGD splenic enzymes, the\nresults were the same except that K(is)app(AJGD)\/K(is)app(normal) = 4-17 for dNM and 1-\ndeoxy-1-amino-\u00df-glucose; this ratio was \u2245 1 with most other glycons, and particularly, nojirimycin\nand castanospermine. The results of these studies indicated that the glycon binding site\nof the normal acid \u00df-glucosidase contains important residues for interaction with the C-2, C-3\nand C-4 hydroxyl groups of \u00df-glucose and a residue with pK(a) = 6.7 which was critical to the\nbinding of amine-containing inhibitors and the hydrolysis of substrates. The findings were\nconsistent with a specific alteration in or near the glycon binding site which results in the\nfunctional abnormalities of the mutant AJGD acid \u00df-glucosidase.<\/jats:p>","DOI":"10.1159\/000469161","type":"journal-article","created":{"date-parts":[[2017,8,11]],"date-time":"2017-08-11T17:22:16Z","timestamp":1502472136000},"page":"173-188","source":"Crossref","is-referenced-by-count":29,"title":["Human Acid \u00df-Glucosidase: Inhibition Studies Using Glucose\nAnalogues and pH Variation to Characterize the Normal and\nGaucher Disease Glycon Binding Sites"],"prefix":"10.1159","author":[{"given":"Karen","family":"Osiecki-Newman","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gunter","family":"Legler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"Grace","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tama","family":"Dinur","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shimon","family":"Gatt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert","family":"Desnick","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gregory","family":"Grabowski","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"127","published-online":{"date-parts":[[2017,8,11]]},"container-title":["Enzyme"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/karger.com\/article-pdf\/doi\/10.1159\/000469161","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T12:47:10Z","timestamp":1777553230000},"score":1,"resource":{"primary":{"URL":"https:\/\/karger.com\/article\/doi\/10.1159\/000469161"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,8,11]]},"references-count":0,"URL":"https:\/\/doi.org\/10.1159\/000469161","archive":["Portico"],"relation":{},"ISSN":["0013-9432","2504-2564"],"issn-type":[{"value":"0013-9432","type":"print"},{"value":"2504-2564","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,8,11]]}}}