Youlin Xia, Tairan Yuwen, Aizhuo Liu, Charalampos G. Kalodimos
{"title":"13CHD2 - 13C-CEST剖面中h -去耦边带的去除","authors":"Youlin Xia, Tairan Yuwen, Aizhuo Liu, Charalampos G. Kalodimos","doi":"10.1007/s10858-021-00362-0","DOIUrl":null,"url":null,"abstract":"<p>A unique aspect of NMR is its capacity to provide integrated insight into both the structure and intrinsic dynamics of biomolecules. Chemical exchange phenomena that often serve as probes of dynamic processes in biological macromolecules can be quantitatively investigated with chemical exchange saturation transfer (CEST) experiments. <sup>2</sup>H-decoupling sidebands, however, always occur in the profiles of <sup>13</sup>CHD<sub>2</sub> <sup>13</sup>C-CEST experiments when using the simple CW (continuous wave) method, which may obscure the detection of minor dips of excited states. Traditionally, these sidebands are manually eliminated from the profiles before data analysis by removing experimental points in the range of <sup>2</sup>H-decoupling field strength?±50?Hz away from the major dips of the ground state on either side of the dips. Unfortunately, this may also eliminate potential minor dips if they overlap with the decoupling sidebands. Here, we developed methods that use pseudo-continuous waves with variable RF amplitudes distributed onto ramps for <sup>2</sup>H decoupling. The new methods were thoroughly validated on Bruker spectrometers at a range of fields (<sup>1</sup>H frequencies of 600, 700, and 850?MHz, and 1.1 GHz). By using these methods, we successfully removed the sidebands from the NMR profiles of <sup>13</sup>CHD<sub>2</sub> <sup>13</sup>C-CEST experiments.</p>","PeriodicalId":613,"journal":{"name":"Journal of Biomolecular NMR","volume":"75 2-3","pages":"133 - 142"},"PeriodicalIF":1.3000,"publicationDate":"2021-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s10858-021-00362-0","citationCount":"3","resultStr":"{\"title\":\"Removal of 2H-decoupling sidebands in 13CHD2 13C-CEST profiles\",\"authors\":\"Youlin Xia, Tairan Yuwen, Aizhuo Liu, Charalampos G. Kalodimos\",\"doi\":\"10.1007/s10858-021-00362-0\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>A unique aspect of NMR is its capacity to provide integrated insight into both the structure and intrinsic dynamics of biomolecules. Chemical exchange phenomena that often serve as probes of dynamic processes in biological macromolecules can be quantitatively investigated with chemical exchange saturation transfer (CEST) experiments. <sup>2</sup>H-decoupling sidebands, however, always occur in the profiles of <sup>13</sup>CHD<sub>2</sub> <sup>13</sup>C-CEST experiments when using the simple CW (continuous wave) method, which may obscure the detection of minor dips of excited states. Traditionally, these sidebands are manually eliminated from the profiles before data analysis by removing experimental points in the range of <sup>2</sup>H-decoupling field strength?±50?Hz away from the major dips of the ground state on either side of the dips. Unfortunately, this may also eliminate potential minor dips if they overlap with the decoupling sidebands. Here, we developed methods that use pseudo-continuous waves with variable RF amplitudes distributed onto ramps for <sup>2</sup>H decoupling. The new methods were thoroughly validated on Bruker spectrometers at a range of fields (<sup>1</sup>H frequencies of 600, 700, and 850?MHz, and 1.1 GHz). By using these methods, we successfully removed the sidebands from the NMR profiles of <sup>13</sup>CHD<sub>2</sub> <sup>13</sup>C-CEST experiments.</p>\",\"PeriodicalId\":613,\"journal\":{\"name\":\"Journal of Biomolecular NMR\",\"volume\":\"75 2-3\",\"pages\":\"133 - 142\"},\"PeriodicalIF\":1.3000,\"publicationDate\":\"2021-03-20\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1007/s10858-021-00362-0\",\"citationCount\":\"3\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Biomolecular NMR\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://link.springer.com/article/10.1007/s10858-021-00362-0\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Biomolecular NMR","FirstCategoryId":"99","ListUrlMain":"https://link.springer.com/article/10.1007/s10858-021-00362-0","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Removal of 2H-decoupling sidebands in 13CHD2 13C-CEST profiles
A unique aspect of NMR is its capacity to provide integrated insight into both the structure and intrinsic dynamics of biomolecules. Chemical exchange phenomena that often serve as probes of dynamic processes in biological macromolecules can be quantitatively investigated with chemical exchange saturation transfer (CEST) experiments. 2H-decoupling sidebands, however, always occur in the profiles of 13CHD213C-CEST experiments when using the simple CW (continuous wave) method, which may obscure the detection of minor dips of excited states. Traditionally, these sidebands are manually eliminated from the profiles before data analysis by removing experimental points in the range of 2H-decoupling field strength?±50?Hz away from the major dips of the ground state on either side of the dips. Unfortunately, this may also eliminate potential minor dips if they overlap with the decoupling sidebands. Here, we developed methods that use pseudo-continuous waves with variable RF amplitudes distributed onto ramps for 2H decoupling. The new methods were thoroughly validated on Bruker spectrometers at a range of fields (1H frequencies of 600, 700, and 850?MHz, and 1.1 GHz). By using these methods, we successfully removed the sidebands from the NMR profiles of 13CHD213C-CEST experiments.
期刊介绍:
The Journal of Biomolecular NMR provides a forum for publishing research on technical developments and innovative applications of nuclear magnetic resonance spectroscopy for the study of structure and dynamic properties of biopolymers in solution, liquid crystals, solids and mixed environments, e.g., attached to membranes. This may include:
Three-dimensional structure determination of biological macromolecules (polypeptides/proteins, DNA, RNA, oligosaccharides) by NMR.
New NMR techniques for studies of biological macromolecules.
Novel approaches to computer-aided automated analysis of multidimensional NMR spectra.
Computational methods for the structural interpretation of NMR data, including structure refinement.
Comparisons of structures determined by NMR with those obtained by other methods, e.g. by diffraction techniques with protein single crystals.
New techniques of sample preparation for NMR experiments (biosynthetic and chemical methods for isotope labeling, preparation of nutrients for biosynthetic isotope labeling, etc.). An NMR characterization of the products must be included.