Magic Angle Spinning NMR Structure Determination of Proteins from Pseudocontact Shifts

Handle URI:
http://hdl.handle.net/10754/598747
Title:
Magic Angle Spinning NMR Structure Determination of Proteins from Pseudocontact Shifts
Authors:
Li, Jianping; Pilla, Kala Bharath; Li, Qingfeng; Zhang, Zhengfeng; Su, Xuncheng; Huber, Thomas; Yang, Jun
Abstract:
Magic angle spinning solid-state NMR is a unique technique to study atomic-resolution structure of biomacromolecules which resist crystallization or are too large to study by solution NMR techniques. However, difficulties in obtaining sufficient number of long-range distance restraints using dipolar coupling based spectra hamper the process of structure determination of proteins in solid-state NMR. In this study it is shown that high-resolution structure of proteins in solid phase can be determined without the use of traditional dipolar-dipolar coupling based distance restraints by combining the measurements of pseudocontact shifts (PCSs) with Rosetta calculations. The PCSs were generated by chelating exogenous paramagnetic metal ions to a tag 4-mercaptomethyl-dipicolinic acid, which is covalently attached to different residue sites in a 56-residue immunoglobulin-binding domain of protein G (GB1). The long-range structural restraints with metal-nucleus distance of up to ∼20 Å are quantitatively extracted from experimentally observed PCSs, and these are in good agreement with the distances back-calculated using an X-ray structure model. Moreover, we demonstrate that using several paramagnetic ions with varied paramagnetic susceptibilities as well as the introduction of paramagnetic labels at different sites can dramatically increase the number of long-range restraints and cover different regions of the protein. The structure generated from solid-state NMR PCSs restraints combined with Rosetta calculations has 0.7 Å root-mean-square deviation relative to X-ray structure. © 2013 American Chemical Society.
Citation:
Li J, Pilla KB, Li Q, Zhang Z, Su X, et al. (2013) Magic Angle Spinning NMR Structure Determination of Proteins from Pseudocontact Shifts. Journal of the American Chemical Society 135: 8294–8303. Available: http://dx.doi.org/10.1021/ja4021149.
Publisher:
American Chemical Society (ACS)
Journal:
Journal of the American Chemical Society
Issue Date:
5-Jun-2013
DOI:
10.1021/ja4021149
PubMed ID:
23646876
Type:
Article
ISSN:
0002-7863; 1520-5126
Sponsors:
J.Y. thanks Tatyana Polenova for valuable discussions. The authors thank Xu Zhang for help in solution NMR experiments. This work is supported by grants from the National Natural Science Foundation of China (21075133, 21173259, 21073101) and the National Basic Research Program of China (2009CB918600). We are thankful to King Abdulla University of Science and Technology (KAUST), Saudi Arabia, for providing access to the Blue Gene/P (Shaheen) supercomputer. T.H. acknowledges funding from the Australian Research Council, including a Future Fellowship (FT0991709) and project grant (DP120100561).
Appears in Collections:
Publications Acknowledging KAUST Support

Full metadata record

DC FieldValue Language
dc.contributor.authorLi, Jianpingen
dc.contributor.authorPilla, Kala Bharathen
dc.contributor.authorLi, Qingfengen
dc.contributor.authorZhang, Zhengfengen
dc.contributor.authorSu, Xunchengen
dc.contributor.authorHuber, Thomasen
dc.contributor.authorYang, Junen
dc.date.accessioned2016-02-25T13:40:24Zen
dc.date.available2016-02-25T13:40:24Zen
dc.date.issued2013-06-05en
dc.identifier.citationLi J, Pilla KB, Li Q, Zhang Z, Su X, et al. (2013) Magic Angle Spinning NMR Structure Determination of Proteins from Pseudocontact Shifts. Journal of the American Chemical Society 135: 8294–8303. Available: http://dx.doi.org/10.1021/ja4021149.en
dc.identifier.issn0002-7863en
dc.identifier.issn1520-5126en
dc.identifier.pmid23646876en
dc.identifier.doi10.1021/ja4021149en
dc.identifier.urihttp://hdl.handle.net/10754/598747en
dc.description.abstractMagic angle spinning solid-state NMR is a unique technique to study atomic-resolution structure of biomacromolecules which resist crystallization or are too large to study by solution NMR techniques. However, difficulties in obtaining sufficient number of long-range distance restraints using dipolar coupling based spectra hamper the process of structure determination of proteins in solid-state NMR. In this study it is shown that high-resolution structure of proteins in solid phase can be determined without the use of traditional dipolar-dipolar coupling based distance restraints by combining the measurements of pseudocontact shifts (PCSs) with Rosetta calculations. The PCSs were generated by chelating exogenous paramagnetic metal ions to a tag 4-mercaptomethyl-dipicolinic acid, which is covalently attached to different residue sites in a 56-residue immunoglobulin-binding domain of protein G (GB1). The long-range structural restraints with metal-nucleus distance of up to ∼20 Å are quantitatively extracted from experimentally observed PCSs, and these are in good agreement with the distances back-calculated using an X-ray structure model. Moreover, we demonstrate that using several paramagnetic ions with varied paramagnetic susceptibilities as well as the introduction of paramagnetic labels at different sites can dramatically increase the number of long-range restraints and cover different regions of the protein. The structure generated from solid-state NMR PCSs restraints combined with Rosetta calculations has 0.7 Å root-mean-square deviation relative to X-ray structure. © 2013 American Chemical Society.en
dc.description.sponsorshipJ.Y. thanks Tatyana Polenova for valuable discussions. The authors thank Xu Zhang for help in solution NMR experiments. This work is supported by grants from the National Natural Science Foundation of China (21075133, 21173259, 21073101) and the National Basic Research Program of China (2009CB918600). We are thankful to King Abdulla University of Science and Technology (KAUST), Saudi Arabia, for providing access to the Blue Gene/P (Shaheen) supercomputer. T.H. acknowledges funding from the Australian Research Council, including a Future Fellowship (FT0991709) and project grant (DP120100561).en
dc.publisherAmerican Chemical Society (ACS)en
dc.titleMagic Angle Spinning NMR Structure Determination of Proteins from Pseudocontact Shiftsen
dc.typeArticleen
dc.identifier.journalJournal of the American Chemical Societyen
dc.contributor.institutionChinese Academy of Sciences, Beijing, Chinaen
dc.contributor.institutionNankai University, Tianjin, Chinaen
dc.contributor.institutionAustralian National University, Canberra, Australiaen

Related articles on PubMed

All Items in KAUST are protected by copyright, with all rights reserved, unless otherwise indicated.