{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,29]],"date-time":"2026-08-29T01:56:50Z","timestamp":1787968610037,"version":"build-2784847793"},"reference-count":30,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2019,4,29]],"date-time":"2019-04-29T00:00:00Z","timestamp":1556496000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010663","name":"H2020 European Research Council","doi-asserted-by":"publisher","award":["ERC advanced grant \/ 290836 CHEMBIOSPHING"],"award-info":[{"award-number":["ERC advanced grant \/ 290836 CHEMBIOSPHING"]}],"id":[{"id":"10.13039\/100010663","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Traffic"],"published-print":{"date-parts":[[2019,5]]},"abstract":"<jats:p>\u03b2\u2010Glucocerebrosidase (GBA) is the enzyme that degrades glucosylceramide in lysosomes. Defects in GBA that result in overall loss of enzymatic activity give rise to the lysosomal storage disorder Gaucher disease, which is characterized by the accumulation of glucosylceramide in tissue macrophages. Gaucher disease is currently treated by infusion of mannose receptor\u2010targeted recombinant GBA. The recombinant GBA is thought to reach the lysosomes of macrophages, based on the impressive clinical response that is observed in Gaucher patients (type 1) receiving this enzyme replacement therapy. In this study, we used cyclophellitol\u2010derived activity\u2010based probes (ABPs) with a fluorescent reporter that irreversibly bind to the catalytic pocket of GBA, to visualize the active enzymes in a correlative microscopy approach. The uptake of pre\u2010labeled recombinant enzyme was monitored by fluorescence and electron microscopy in human fibroblasts that stably expressed the mannose receptor. The endogenous active enzyme was simultaneously visualized by in situ labeling with the ABP containing an orthogonal fluorophore. This method revealed the efficient delivery of recombinant GBA to lysosomal target compartments that contained endogenous active enzyme.<\/jats:p>\n                  <jats:p>\n                    <jats:inline-graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"graphic\/tra12641-gra-0001.png\" xlink:title=\"image\"\/>\n                  <\/jats:p>","DOI":"10.1111\/tra.12641","type":"journal-article","created":{"date-parts":[[2019,3,21]],"date-time":"2019-03-21T04:44:20Z","timestamp":1553143460000},"page":"346-356","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Localization of active endogenous and exogenous \u03b2\u2010glucocerebrosidase by correlative light\u2010electron microscopy in human fibroblasts"],"prefix":"10.1111","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7633-0186","authenticated-orcid":false,"given":"Eline","family":"van Meel","sequence":"first","affiliation":[{"name":"Department of Medical Biochemistry, Leiden Institute of Chemistry Leiden University  Leiden the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erik","family":"Bos","sequence":"additional","affiliation":[{"name":"Department of Cell and Chemical Biology Leiden University Medical Center  Leiden the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martijn J. 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