Research Article

Magic angle spinning NMR spectroscopy of thioredoxin reassemblies

  1. Jun Yang1,
  2. Sivakumar Paramasivam1,
  3. Dabeiba Marulanda1,†,
  4. Marcela Cataldi2,
  5. Maria Luisa Tasayco2,
  6. Tatyana Polenova1,*

Article first published online: 21 DEC 2007

DOI: 10.1002/mrc.2092

Issue

Magnetic Resonance in Chemistry

Magnetic Resonance in Chemistry

Special Issue: New techniques in solid-state NMR

Volume 45, Issue S1, pages S73–S83, December 2007

Additional Information

How to Cite

Yang, J., Paramasivam, S., Marulanda, D., Cataldi, M., Tasayco, M. L. and Polenova, T. (2007), Magic angle spinning NMR spectroscopy of thioredoxin reassemblies. Magn. Reson. Chem., 45: S73–S83. doi: 10.1002/mrc.2092

Author Information

  1. 1

    Department of Chemistry and Biochemistry, University of Delaware, Newark, DE 19716, USA

  2. 2

    Department of Chemistry, The City College of New York, Convent Avenue at 138th Street, New York, NY 10031, USA

  1. †

    Currently on leave from the New York Medical College

*Department of Chemistry and Biochemistry, University of Delaware, Newark, DE 19716, USA.

Publication History

  1. Issue published online: 21 DEC 2007
  2. Article first published online: 21 DEC 2007
  3. Manuscript Accepted: 23 AUG 2007
  4. Manuscript Revised: 20 AUG 2007
  5. Manuscript Received: 18 JUN 2007

Funded by

  • National Science Foundation. Grant Numbers: NSF-CAREER CHE-0237612, MCB-0517592
  • National Institutes of Health. Grant Numbers: P20-17716, 5G12RR03060, GM66354

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Keywords:

  • solid-state NMR;
  • resonance assignments;
  • magic angle spinning;
  • 13C;
  • 15N;
  • thioredoxin;
  • reassembled thioredoxin;
  • secondary structure;
  • tertiary constraints

Abstract

Differentially isotopically enriched 1–73(13C,15N)/74–108(15N) and 1–73(15N)/74–108(13C,15N) Escherichia coli thioredoxin reassemblies prepared by fragment complementation were investigated by high-resolution magic angle spinning solid-state NMR spectroscopy. Nearly complete resonance assignments, secondary and tertiary structure analysis are reported for 1–73(13C,15N)/74–108(15N) reassembled thioredoxin. Temperature dependence of the dipolar-assisted rotational resonance (DARR) spectra reveals the residues undergoing intermediate timescale motions at temperatures below − 15 °C. Analysis of the DARR intensity buildups as a function of mixing time in these reassemblies indicates that at long mixing times medium- and long-range cross-peaks do not experience dipolar truncation, suggesting that isotopic dilution is not required for gaining nontrivial distance restraints for structure calculations. Copyright © 2007 John Wiley & Sons, Ltd.

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