Figshare最新文献

{"title":"Leveraging High-ResolutionnIon Mobility-Mass Spectrometrynfor Cyclic Peptide Soft Spot Identification","authors":"Maria Fawaz (2026360), Congliang Sun (4822185), Yu Feng (445691), Athanasia Qirjollari (18474581), Hubert Josien (1436686), Daniel DeBord (10326810), Ashli Simone (19111416), David L. Williamson (12066880), Kara Pearson (6996275), Raymond J. Gonzalez (18474584), Lisa Vasicek (1785325), Mark T. Cancilla (1961914), Weixun Wang (2057191), Daniel S. Spellman (2649097), Komal Kedia (4173742)","doi":"10.1021/jasms.4c00109.s001","DOIUrl":"https://doi.org/10.1021/jasms.4c00109.s001","url":null,"abstract":"Cyclic peptides are an important class of molecules thatngainednsignificant attention in the field of drug discovery due to theirnunique pharmacological characteristics and enhanced proteolytic stability.nYet, gastrointestinal degradation remains a major hurdle in the discoverynof orally bioavailable cyclic peptides. Soft spot identification (SSID)nof the regions in the cyclic peptide sequence susceptible to amidenhydrolysis by proteases is used in the discovery stage to guide medicinalnchemistry design. SSID can be an arduous task, traditionally performednusing liquid chromatography–tandem mass spectrometry (LC-MS/MS),noften resulting in complex and time-consuming manual analysis, particularlynwhen isomeric linear peptide metabolites chromatographically coelute.nHere, we present an alternative orthogonal approach that entails anhigh-resolution ion mobility (HRIM) system based on Structures fornLossless Ion Manipulation (SLIM) technology interfaced with quadrupolentime-of-flight (QTOF) mass spectrometry to address some of the challengesnassociated with SSID. Two strategies were used to resolve linear isomericnpeptide metabolites: labeled and label-free, both utilizing the HRIMnplatform. The label-free strategy leverages negative polarity to ionizenthe isomers which achieves better separation of the gas phase ionsnin the ion mobility (IM) dimension as compared to positive polarity,nwhich is a more conventional approach when studying proteins and peptides.nThe second approach uses an isotope-labeled dimethyl tag on the terminalnamine group, acting as a “shift reagent” to influencenthe mobility of isomers in the positive mode. This method resultednin baseline separation for the isomers of interest and produced uniquenproduct ions in the fragmentation spectra for unambiguous soft spotnidentification. Both label-free and labeled strategies demonstratednthe ability to solve the challenges associated with SSID for cyclicnpeptides.","PeriodicalId":522907,"journal":{"name":"Figshare","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://figshare.com/articles/journal_contribution/Leveraging_High-Resolution_Ion_Mobility-Mass_Spectrometry_for_Cyclic_Peptide_Soft_Spot_Identification/26265852","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147905940","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}