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ECA biosynthesis begins on the cytoplasmic side of the inner membrane (IM) where glycosyltransferases sequentially add sugar moieties to form a complete repeat unit which is then translocated across the IM by WzxE before being polymerized into short linear chains by WzyE\/WzzE. Research into WecG, the enzyme responsible for generating ECA lipid\u2010II, has not progressed beyond Barr et al. (1988) who described WecG as a membrane protein. Here we revise our understanding of WecG and re\u2010characterize it as a peripherally associated membrane protein. Through the use of Western immunoblotting we show that WecG in <jats:italic>Shigella flexneri<\/jats:italic> is maintained to the IM via its three C\u2010terminal helices and further identify key residues in helix II which are critical for this interaction which has allowed us to identify WecG as a GT\u2010E glycosyltransferase. We investigate the possibility of protein complexes and ultimately show that ECA lipid\u2010I maintains WecG to the membrane which is crucial for its function. This research is the first since Barr et al. (1988) to investigate the biochemistry of WecG and reveals possible novel drug targets to inhibit WecG and thus ECA function and cell viability.<\/jats:p>","DOI":"10.1111\/mmi.14973","type":"journal-article","created":{"date-parts":[[2022,8,12]],"date-time":"2022-08-12T16:39:29Z","timestamp":1660322369000},"page":"403-416","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Subcellular localization of the enterobacterial common antigen <scp>GT\u2010E\u2010<\/scp>like glycosyltransferase, <scp>WecG<\/scp>"],"prefix":"10.1111","volume":"118","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7638-2613","authenticated-orcid":false,"given":"Nicholas","family":"Maczuga","sequence":"first","affiliation":[{"name":"Department of Molecular and Biomedical Sciences, Research Centre for Infectious Diseases, School of Biological Sciences The University of Adelaide  Adelaide South Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth N. H.","family":"Tran","sequence":"additional","affiliation":[{"name":"Department of Molecular and Biomedical Sciences, Research Centre for Infectious Diseases, School of Biological Sciences The University of Adelaide  Adelaide South Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renato","family":"Morona","sequence":"additional","affiliation":[{"name":"Department of Molecular and Biomedical Sciences, Research Centre for Infectious Diseases, School of Biological Sciences The University of Adelaide  Adelaide South Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2022,8,25]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00676-08"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.000701"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/msb4100050"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.170.1.228-233.1988"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1110\/ps.0213502"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00839-10"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.180.22.5875-5884.1998"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.120163297"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkv332"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1021\/acschembio.0c00983"},{"key":"e_1_2_11_12_1","first-page":"1003","article-title":"The 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