Towards single-molecule observation of protein synthesis

David Dulin (1) , Antoine Le Gall (1) , Philippe Bouyer (2) , Karen Perronet (1) , Nathalie Westbrook (1) , Nicolas Soler (3) , Dominique Fourmy (3) , Satoko Yoshizawa (3)

1 Laboratoire Charles Fabry de l'Institut d'Optique / Otique atomique - bio
2 laboratoire Charles Fabry de l'Institut d'Optique / Optique atomique
3 ICSN - Institut de Chimie des Substances Naturelles

The ribosome is the molecular motor responsible for the protein synthesis within all cells. Ribosome motions along the messenger RNA (mRNA) to read the genetic code are asynchronous and occur along multiple kinetic paths. Consequently, a study at the single macromolecule level is desirable to unravel the complex dynamics in- volved. In this communication, we present the development of an advanced surface chemistry to attach an active ribosome to the microscope coverslip and follow the amino-acid incorporation by fluorescence microscopy. The ribo- some is labeled with a quantum dot (QD) in order to localize it on the surface while a specific amino acid (lysine) is marked with Bodipy-FL. This fluorescent dye is small enough to enter the ribosomal channel thus leaving intact ribosomal activity. It should then be possible to observe the protein synthesis in real time as the labeled amino acids are incorporated into the polypeptide chain.

Mots clés: Single molecule; Fluorescence microscopy

Domaines: Biophysique [physics.bio-ph]

David Dulin, Antoine Le Gall, Philippe Bouyer, Karen Perronet, Nathalie Westbrook, et al.. Towards single-molecule observation of protein synthesis. Tecnolaser 2009, Apr 2009, La Havane, Cuba. pp.9. ⟨hal-00546717⟩