Sahil Ahlawat, Subbarao Mohana Venkata Mopidevi, Pravin P. Taware, Sreejith Raran-Kurussi, Kaustubh R. Mote, Vipin Agarwal
{"title":"芳香族侧链自旋的分配及其在快速MAS下距离限制的表征","authors":"Sahil Ahlawat, Subbarao Mohana Venkata Mopidevi, Pravin P. Taware, Sreejith Raran-Kurussi, Kaustubh R. Mote, Vipin Agarwal","doi":"10.1016/j.yjsbx.2022.100082","DOIUrl":null,"url":null,"abstract":"<div><p>The assignment of aromatic side-chain spins has always been more challenging than assigning backbone and aliphatic spins. Selective labeling combined with mutagenesis has been the approach for assigning aromatic spins. This manuscript reports a method for assigning aromatic spins in a fully protonated protein by connecting them to the backbone atoms using a low-power TOBSY sequence. The pulse sequence employs residual polarization and sequential acquisitions techniques to record H<sup>N</sup>- and H<sup>C</sup>-detected spectra in a single experiment. The unambiguous assignment of aromatic spins also enables the characterization of <sup>1</sup>H–<sup>1</sup>H distance restraints involving aromatic spins. Broadband (RFDR) and selective (BASS-SD) recoupling sequences were used to generate H<sup>N</sup>-Η<sup>C</sup>, H<sup>C</sup>-H<sup>N</sup> and H<sup>C</sup>-H<sup>C</sup> restraints involving the side-chain proton spins of aromatic residues. This approach has been demonstrated on a fully protonated U-[<sup>13</sup>C,<sup>15</sup>N] labeled GB1 sample at 95–100 kHz MAS.</p></div>","PeriodicalId":17238,"journal":{"name":"Journal of Structural Biology: X","volume":null,"pages":null},"PeriodicalIF":3.5000,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/c8/90/main.PMC9817166.pdf","citationCount":"2","resultStr":"{\"title\":\"Assignment of aromatic side-chain spins and characterization of their distance restraints at fast MAS\",\"authors\":\"Sahil Ahlawat, Subbarao Mohana Venkata Mopidevi, Pravin P. Taware, Sreejith Raran-Kurussi, Kaustubh R. Mote, Vipin Agarwal\",\"doi\":\"10.1016/j.yjsbx.2022.100082\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>The assignment of aromatic side-chain spins has always been more challenging than assigning backbone and aliphatic spins. Selective labeling combined with mutagenesis has been the approach for assigning aromatic spins. This manuscript reports a method for assigning aromatic spins in a fully protonated protein by connecting them to the backbone atoms using a low-power TOBSY sequence. The pulse sequence employs residual polarization and sequential acquisitions techniques to record H<sup>N</sup>- and H<sup>C</sup>-detected spectra in a single experiment. The unambiguous assignment of aromatic spins also enables the characterization of <sup>1</sup>H–<sup>1</sup>H distance restraints involving aromatic spins. Broadband (RFDR) and selective (BASS-SD) recoupling sequences were used to generate H<sup>N</sup>-Η<sup>C</sup>, H<sup>C</sup>-H<sup>N</sup> and H<sup>C</sup>-H<sup>C</sup> restraints involving the side-chain proton spins of aromatic residues. This approach has been demonstrated on a fully protonated U-[<sup>13</sup>C,<sup>15</sup>N] labeled GB1 sample at 95–100 kHz MAS.</p></div>\",\"PeriodicalId\":17238,\"journal\":{\"name\":\"Journal of Structural Biology: X\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.5000,\"publicationDate\":\"2023-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/c8/90/main.PMC9817166.pdf\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Structural Biology: X\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S259015242200023X\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Structural Biology: X","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S259015242200023X","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Assignment of aromatic side-chain spins and characterization of their distance restraints at fast MAS
The assignment of aromatic side-chain spins has always been more challenging than assigning backbone and aliphatic spins. Selective labeling combined with mutagenesis has been the approach for assigning aromatic spins. This manuscript reports a method for assigning aromatic spins in a fully protonated protein by connecting them to the backbone atoms using a low-power TOBSY sequence. The pulse sequence employs residual polarization and sequential acquisitions techniques to record HN- and HC-detected spectra in a single experiment. The unambiguous assignment of aromatic spins also enables the characterization of 1H–1H distance restraints involving aromatic spins. Broadband (RFDR) and selective (BASS-SD) recoupling sequences were used to generate HN-ΗC, HC-HN and HC-HC restraints involving the side-chain proton spins of aromatic residues. This approach has been demonstrated on a fully protonated U-[13C,15N] labeled GB1 sample at 95–100 kHz MAS.
京公网安备 11010802042870号
京ICP备2023020795号-1
分享
求助内容：
应助结果提醒方式：
扫码关注我们
