Next Article in Journal
Mulberry Fruit Extract Protects Pancreatic β-Cells against Hydrogen Peroxide-Induced Apoptosis via Antioxidative Activity
Next Article in Special Issue
Cytotoxic Activity of 3,6-Dihydroxyflavone in Human Cervical Cancer Cells and Its Therapeutic Effect on c-Jun N-Terminal Kinase Inhibition
Previous Article in Journal
A New Xanthone from Moutabea guianensis Aubl
Article Menu

Export Article

Open AccessCommunication
Molecules 2014, 19(7), 8890-8903; doi:10.3390/molecules19078890

Using MUSIC and CC(CO)NH for Backbone Assignment of Two Medium-Sized Proteins Not Fully Accessible to Standard 3D NMR

1
Department of Chemistry, University of Bergen, PO Box 7800, 5020 Bergen, Norway
2
Department of Clinical Science, University of Bergen, PO Box 7804, 5020 Bergen, Norway
*
Author to whom correspondence should be addressed.
Received: 5 May 2014 / Revised: 5 June 2014 / Accepted: 12 June 2014 / Published: 26 June 2014
(This article belongs to the Special Issue Design and Study of Kinase Inhibitors)
View Full-Text   |   Download PDF [306 KB, uploaded 26 June 2014]   |  

Abstract

The backbone assignment of medium-sized proteins is rarely as straightforward as that of small proteins, and thus often requires creative solutions. Here, we describe the application of a combination of standard 3D heteronuclear methods with CC(CO)NH and a variety of MUltiplicity Selective In-phase Coherence transfer (MUSIC) experiments. Both CC(CO)NH and MUSIC are, in theory, very powerful methods for the backbone assignment of proteins. Due to low sensitivity, their use has usually been linked to small proteins only. However, we found that combining CC(CO)NH and MUSIC experiments simplified the assignment of two challenging medium-sized proteins of 13 and 19.5 kDa, respectively. These methods are to some extent complementary to each other: CC(CO)NH acquired with a long isotropic mixing time can identify amino acids with large aliphatic side chains. Whereas the most sensitive MUSIC experiments identify amino acid types that cannot be detected by CC(CO)NH, comprising the residues with acid and amide groups, and aromatic rings in their side chains. Together these methods provide a means of identifying the majority of peaks in the 2D 15N HSQC spectrum which simplifies the backbone assignment work even for proteins, e.g., small kinases, whose standard spectra resulted in little spectral resolution and low signal intensities. View Full-Text
Keywords: protein NMR spectroscopy; backbone assignment; CC(CO)NH; MUSIC protein NMR spectroscopy; backbone assignment; CC(CO)NH; MUSIC
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Brenner, A.K.; Frøystein, N.Å. Using MUSIC and CC(CO)NH for Backbone Assignment of Two Medium-Sized Proteins Not Fully Accessible to Standard 3D NMR. Molecules 2014, 19, 8890-8903.

Show more citation formats Show less citations formats

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]

Molecules EISSN 1420-3049 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top