Footprinting of protein interactions by tritium labeling

Guillaume Mousseau; Quentin Raffy; Olivier P Thomas; Morgane Agez; Robert Thai; Jean Philippe Renault; Serge Pin; Françoise Ochsenbein; Jean-Christophe Cintrat; Bernard Rousseau

doi:10.1021/bi100031a

Footprinting of protein interactions by tritium labeling

Biochemistry. 2010 May 25;49(20):4297-9. doi: 10.1021/bi100031a.

Authors

Guillaume Mousseau¹, Quentin Raffy, Olivier P Thomas, Morgane Agez, Robert Thai, Jean Philippe Renault, Serge Pin, Françoise Ochsenbein, Jean-Christophe Cintrat, Bernard Rousseau

Affiliation

¹ IBITECS, CNRS CEA, F-91191 Gif-sur-Yvette, France.

PMID: 20415454
DOI: 10.1021/bi100031a

Abstract

A new footprinting method for mapping protein interactions has been developed, using tritium as a radioactive label. As residues involved in an interaction are less labeled when the complex is formed, they can be identified via comparison of the tritium incorporation of each residue of the bound protein with that of the unbound one. Application of this footprinting method to the complex formed by the histone H3 fragment H3(122-135) and the protein hAsf1A(1-156) afforded data in good agreement with NMR results.

Publication types

Validation Study

MeSH terms

Amino Acid Sequence
Efficiency
Histones / chemistry
Histones / metabolism
Humans
Isotope Labeling / methods*
Models, Biological
Nuclear Magnetic Resonance, Biomolecular
Nucleosomes / chemistry
Nucleosomes / metabolism
Peptide Fragments / chemistry
Peptide Fragments / metabolism
Protein Binding
Protein Footprinting / methods*
Protein Interaction Mapping / methods
Proteins / chemistry
Proteins / metabolism*
Sensitivity and Specificity
Tritium

Substances

Histones
Nucleosomes
Peptide Fragments
Proteins
Tritium