{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T22:17:59Z","timestamp":1772835479967,"version":"3.50.1"},"reference-count":30,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2020,12,2]],"date-time":"2020-12-02T00:00:00Z","timestamp":1606867200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100012806","name":"High Point University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100012806","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["FEBS Open Bio"],"published-print":{"date-parts":[[2021,1]]},"abstract":"<jats:p>The calcitonin receptor (CTR) has a large extracellular domain (ECD) with multiple <jats:italic>N<\/jats:italic>\u2010glycosylation sites. An asparagine (Asn)\u2010linked <jats:italic>N<\/jats:italic>\u2010acetylglucosamine (GlcNAc) of CTR ECD N130 was previously reported to enhance peptide hormone binding affinity for CTR ECD. CTR forms a complex with an accessory protein RAMP, and the RAMP:CTR complex gains affinity for peptide hormone amylin as the amylin receptor (AMY). Although <jats:italic>N<\/jats:italic>\u2010glycosylation of AMY ECD was reported to enhance peptide hormone affinity, it remains underexplored which <jats:italic>N<\/jats:italic>\u2010glycosites of AMY ECD are responsible for peptide affinity enhancement and it is unclear whether an Asn\u2010linked GlcNAc of the <jats:italic>N<\/jats:italic>\u2010glycosites plays a critical role. Here, I investigated the role of the Asn\u2010linked GlcNAc of CTR N130 in the affinity of an antagonistic amylin analog (AC413) for AMY<jats:sub>2<\/jats:sub> ECD (the RAMP2 ECD:CTR ECD complex). I used Endo H\u2010treated CTR ECD in which <jats:italic>N<\/jats:italic>\u2010glycans were trimmed to an Asn\u2010linked GlcNAc on each of the <jats:italic>N<\/jats:italic>\u2010glycosites. I incubated Endo H\u2010treated CTR ECD with excess of glycan\u2010free RAMP2 ECD to produce the RAMP2 ECD:CTR ECD complex. Using this coincubation system, I found that the RAMP2 ECD complex with Endo H\u2010treated CTR ECD with N130D mutation showed a fourfold decrease in AC413 affinity compared with the RAMP2 ECD complex with Endo H\u2010treated CTR ECD WT. In contrast, RAMP2 ECD <jats:italic>N<\/jats:italic>\u2010glycosylation did not affect peptide binding affinity. These results indicate that the Asn\u2010linked GlcNAc of CTR N130 is an important peptide affinity enhancer for AMY<jats:sub>2<\/jats:sub> ECD and reveals a significant role of the Asn\u2010linked GlcNAc in AMY<jats:sub>2<\/jats:sub> function.<\/jats:p>","DOI":"10.1002\/2211-5463.13042","type":"journal-article","created":{"date-parts":[[2020,11,24]],"date-time":"2020-11-24T05:25:30Z","timestamp":1606195530000},"page":"195-206","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Asn\u2010linked N\u2013acetylglucosamine of the amylin receptor 2 extracellular domain enhances peptide ligand affinity"],"prefix":"10.1002","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7262-159X","authenticated-orcid":false,"given":"Sangmin","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Basic Pharmaceutical Sciences Fred Wilson School of Pharmacy High Point University High Point NC 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