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Sci Rep. 2017 Dec 1;7(1):16696. doi: 10.1038/s41598-017-16959-1.

The repeat region of cortactin is intrinsically disordered in solution.

Author information

1
Department of Pharmacology, Yale University School of Medicine, New Haven, CT, 06520, USA.
2
Department of Pathology, Yale University School of Medicine, New Haven, CT, 06520, USA.
3
Department of Chemistry, Florida State University, 600 W., College Avenue, Tallahassee, FL, 32306, USA.
4
Biopharmaceutical Analytical Sciences, Biopharm R&D, GlaxoSmithKline, 709 Swedeland Road, King of Prussia, PA, 19406, USA.
5
Department of Cell Biology, Yale University School of Medicine, New Haven, CT, 06520, USA.
6
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, 06520, USA.
7
Yale MS & Proteomics Resource, Yale University, New Haven, CT, 06520, USA.
8
Ion Cyclotron Resonance Program, National High Magnetic Field Laboratory, 1800 E. Paul Dirac Dr., Tallahassee, FL, 32310, USA.
9
Department of Pharmacology, Yale University School of Medicine, New Haven, CT, 06520, USA. titus.boggon@yale.edu.
10
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, 06520, USA. titus.boggon@yale.edu.

Abstract

The multi-domain protein, cortactin, contains a 37-residue repeating motif that binds to actin filaments. This cortactin repeat region comprises 6½ similar copies of the motif and binds actin filaments. To better understand this region of cortactin, and its fold, we conducted extensive biophysical analysis. Size exclusion chromatography with multi-angle light scattering (SEC-MALS) reveals that neither constructs of the cortactin repeats alone or together with the adjacent helical region homo-oligomerize. Using circular dichroism (CD) we find that in solution the cortactin repeats resemble a coil-like intrinsically disordered protein. Small-angle X-ray scattering (SAXS) also indicates that the cortactin repeats are intrinsically unfolded, and the experimentally observed radius of gyration (R g) is coincidental to that calculated by the program Flexible-Meccano for an unfolded peptide of this length. Finally, hydrogen-deuterium exchange mass spectrometry (HDX-MS) indicates that the domain contains limited hydrophobic core regions. These experiments therefore provide evidence that in solution the cortactin repeat region of cortactin is intrinsically disordered.

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