Footprinting of protein interactions by tritium labeling

Guillaume Mousseau (1) , Quentin Raffy (2) , Olivier Thomas (3, 1, 4, 5) , Morgane Agez (6, 1) , Robert Thai (7) , Jean Philippe Renault (2) , Serge Pin (2) , Françoise Ochsenbein (1, 8) , Jean-Christophe Cintrat (9) , Bernard Rousseau (9)

1 IBITECS - Institut de Biologie et de Technologies de Saclay
2 SIS2M UMR 3299 - Service Interdisciplinaire sur les Systèmes Moléculaires et les Matériaux (ex SCM)
3 LERES - Laboratoire d'étude et de recherche en environnement et santé
4 EHESP - École des Hautes Études en Santé Publique [EHESP]
5 IRSEA - Institut de Recherche en Sémiochimie et Ethologie Appliquée
6 IGBMC - Institut de Génétique et de Biologie Moléculaire et Cellulaire
7 SIMoS - Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO)
8 SMPSD - Système membranaires, photobiologie, stress et détoxication
9 SCBM - Service de Chimie Bio-Organique et de Marquage

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 H3122−135 and the protein hAsf1A1−156 afforded data in good agreement with NMR results.

Keywords: Hydrogen isotopes; Labeling; Nuclear magnetic resonance spectroscopy; Peptides and proteins

Domains: Analytical chemistry; Radiochemistry; Biochemistry [q-bio.BM]; Structural Biology [q-bio.BM]; Pharmacology; Santé publique et épidémiologie

Guillaume Mousseau, Quentin Raffy, Olivier Thomas, Morgane Agez, Robert Thai, et al.. Footprinting of protein interactions by tritium labeling. British Journal of Pharmacology, 2010, 159 (2), pp.316-325. ⟨10.1021/bi100031a⟩. ⟨cea-02000386⟩