Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016 Sep;66(1):21-35.
doi: 10.1007/s10858-016-0054-9. Epub 2016 Sep 1.

Resonance assignment of disordered protein with repetitive and overlapping sequence using combinatorial approach reveals initial structural propensities and local restrictions in the denatured state

Nikita Malik  1 , Ashutosh Kumar  2
Affiliations

Resonance assignment of disordered protein with repetitive and overlapping sequence using combinatorial approach reveals initial structural propensities and local restrictions in the denatured state

Nikita Malik et al. J Biomol NMR. 2016 Sep.

Abstract

NMR resonance assignment of intrinsically disordered proteins poses a challenge because of the limited dispersion of amide proton chemical shifts. This becomes even more complex with the increase in the size of the system. Residue specific selective labeling/unlabeling experiments have been used to resolve the overlap, but require multiple sample preparations. Here, we demonstrate an assignment strategy requiring only a single sample of uniformly labeled (13)C,(15)N-protein. We have used a combinatorial approach, involving 3D-HNN, CC(CO)NH and 2D-MUSIC, which allowed us to assign a denatured centromeric protein Cse4 of 229 residues. Further, we show that even the less sensitive experiments, when used in an efficient manner can lead to the complete assignment of a complex system without the use of specialized probes in a relatively short time frame. The assignment of the amino acids discloses the presence of local structural propensities even in the denatured state accompanied by restricted motion in certain regions that provides insights into the early folding events of the protein.

Keywords: Centromeric protein; Denatured protein; Intrinsically disordered protein; MUSIC; Protein dynamics.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

References

    1. Biochemistry. 1996 Feb 27;35(8):2674-86 - PubMed
    1. J Magn Reson. 2004 Sep;170(1):15-21 - PubMed
    1. Nature. 2005 Oct 13;437(7061):1053-6 - PubMed
    1. Methods Enzymol. 1989;177:44-73 - PubMed
    1. Nature. 1997 Sep 18;389(6648):251-60 - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources