{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T02:18:52Z","timestamp":1791425932194,"version":"4.3.3"},"reference-count":0,"publisher":"American Society for Microbiology","issue":"3","license":[{"start":{"date-parts":[[1993,3,1]],"date-time":"1993-03-01T00:00:00Z","timestamp":730944000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[1993,3]]},"abstract":"<jats:p>The primary amine coupling reagents succinimidyl-6-biotinamido-hexanoate (NHS-A-biotin) and sulfosuccinimidyl-6-biotinamido-hexanoate (NHS-LC-biotin) were tested for their ability to selectively label Escherichia coli cell envelope proteins in vivo. Probe localization was determined by examining membrane, periplasmic, and cytosolic protein fractions. Both hydrophobic NHS-A-biotin and hydrophilic NHS-LC-biotin were shown to preferentially label outer membrane, periplasmic, and inner membrane proteins. NHS-A- and NHS-LC-biotin were also shown to label a specific inner membrane marker protein (Tet-LacZ). Both probes, however, failed to label a cytosolic marker (the omega fragment of beta-galactosidase). The labeling procedure was also used to label E. coli cells grown in low-salt Luria broth medium supplemented with 0, 10, and 20% sucrose. Outer membrane protein A (OmpA) and OmpC were labeled by both NHS-A- and NHS-LC-biotin at all three sucrose concentrations. In contrast, OmpF was labeled by NHS-A-biotin but not by NHS-LC-biotin in media containing 0 and 10% sucrose. OmpF was not labeled by either NHS-A- or NHS-LC-biotin in E. coli cells grown in medium containing 20% sucrose. Coomassie-stained gels, however, revealed similar quantities of OmpF in E. coli cells grown at all three sucrose concentrations. These data indicate that there was a change in outer membrane structure due to increased osmolarity, which limits accessibility of NHS-A-biotin to OmpF.<\/jats:p>","DOI":"10.1128\/aem.59.3.663-668.1993","type":"journal-article","created":{"date-parts":[[2020,1,1]],"date-time":"2020-01-01T04:46:46Z","timestamp":1577854006000},"page":"663-668","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":25,"title":["In vivo labeling of Escherichia coli cell envelope proteins with N-hydroxysuccinimide esters of biotin"],"prefix":"10.1128","volume":"59","author":[{"given":"J A","family":"Bradburne","sequence":"first","affiliation":[{"name":"School of Biology, Georgia Institute of Technology, Atlanta 30332."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P","family":"Godfrey","sequence":"additional","affiliation":[{"name":"School of Biology, Georgia Institute of Technology, Atlanta 30332."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J H","family":"Choi","sequence":"additional","affiliation":[{"name":"School of Biology, Georgia Institute of Technology, Atlanta 30332."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J N","family":"Mathis","sequence":"additional","affiliation":[{"name":"School of Biology, Georgia Institute of Technology, Atlanta 30332."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aem.59.3.663-668.1993","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aem.59.3.663-668.1993","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T20:54:25Z","timestamp":1645649665000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/aem.59.3.663-668.1993"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1993,3]]},"references-count":0,"journal-issue":{"issue":"3","published-print":{"date-parts":[[1993,3]]}},"alternative-id":["10.1128\/aem.59.3.663-668.1993"],"URL":"https:\/\/doi.org\/10.1128\/aem.59.3.663-668.1993","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[1993,3]]},"assertion":[{"value":"1993-03-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}