Triple resonance 15N NMR relaxation experiments for studies of intrinsically disordered proteins

Triple resonance 15N NMR relaxation experiments for studies of intrinsically disordered proteins Description of protein dynamics is known to be essential in understanding their function. Studies based on a well established $$^{15}\hbox {N}$$ 15 N NMR relaxation methodology have been applied to a large number of systems. However, the low dispersion of $$^{1}\hbox {H}$$ 1 H chemical shifts very often observed within intrinsically disordered proteins complicates utilization of standard 2D HN correlated spectra because a limited number of amino acids can be characterized. Here we present a suite of triple resonance HNCO-type NMR experiments for measurements of five $$^{15}\hbox {N}$$ 15 N relaxation parameters ( $$R_1$$ R 1 , $$R_2$$ R 2 , NOE, cross-correlated relaxation rates $$\Gamma _x$$ Γ x and $$\Gamma _z$$ Γ z ) in doubly $$^{13}\hbox {C}$$ 13 C , $$^{15}\hbox {N}$$ 15 N -labeled proteins. We show that the third spectral dimension combined with non-uniform sampling provides relaxation rates for almost all residues of a protein with extremely poor chemical shift dispersion, the C terminal domain of $$\delta$$ δ -subunit of RNA polymerase from Bacillus subtilis. Comparison with data obtained using a sample labeled by $$^{15}\hbox {N}$$ 15 N only showed that the presence of $$^{13}\hbox {C}$$ 13 C has a negligible effect on $$\Gamma _x$$ Γ x , $$\Gamma _z$$ Γ z , and on the cross-relaxation rate (calculated from NOE and $$R_1$$ R 1 ), and that these relaxation rates can be used to calculate accurate spectral density values. Partially $$^{13}\hbox {C}$$ 13 C -labeled sample was used to test if the observed increase of $$^{15}\hbox {N}$$ 15 N $$R_1$$ R 1 in the presence of $$^{13}\hbox {C}$$ 13 C corresponds to the $$^{15}\hbox {N}-^{13}\hbox {C}$$ 15 N - 13 C dipole–dipole interactions in the $$^{13}\hbox {C}$$ 13 C , $$^{15}\hbox {N}$$ 15 N -labeled sample. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Biomolecular NMR Springer Journals

Triple resonance 15N NMR relaxation experiments for studies of intrinsically disordered proteins

Loading next page...
 
/lp/springer_journal/triple-resonance-15-hbox-n-15-n-nmr-relaxation-experiments-for-studies-oKkDJTKwdQ
Publisher
Springer Netherlands
Copyright
Copyright © 2017 by Springer Science+Business Media B.V.
Subject
Physics; Biological and Medical Physics, Biophysics; Biochemistry, general; Spectroscopy/Spectrometry
ISSN
0925-2738
eISSN
1573-5001
D.O.I.
10.1007/s10858-017-0138-1
Publisher site
See Article on Publisher Site

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 12 million articles from more than
10,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Unlimited reading

Read as many articles as you need. Full articles with original layout, charts and figures. Read online, from anywhere.

Stay up to date

Keep up with your field with Personalized Recommendations and Follow Journals to get automatic updates.

Organize your research

It’s easy to organize your research with our built-in tools.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

Monthly Plan

  • Read unlimited articles
  • Personalized recommendations
  • No expiration
  • Print 20 pages per month
  • 20% off on PDF purchases
  • Organize your research
  • Get updates on your journals and topic searches

$49/month

Start Free Trial

14-day Free Trial

Best Deal — 39% off

Annual Plan

  • All the features of the Professional Plan, but for 39% off!
  • Billed annually
  • No expiration
  • For the normal price of 10 articles elsewhere, you get one full year of unlimited access to articles.

$588

$360/year

billed annually
Start Free Trial

14-day Free Trial