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FRENCH INFRASTRUCTURE FOR INTEGRATED STRUCTURAL BIOLOGY

Mammalian-expression.jpg

INSTRUCT CENTER - FRANCE 1 - IGBMC

ANALYTICAL ULTRA CENTRIFUGATION

Analytical ultracentrifugation (AUC) is a powerful technique for the characterisation of macromolecules and macromolecular self- and hetero-association processes in solution. Two types of complementary experiments, sedimentation velocity and sedimentation equilibrium, can be performed in an analytical ultracentrifuge which is a high-speed centrifuge equipped with an optical detection system. The observation of macromolecules or macromolecular complexes sedimentation gives access to their hydrodynamic and thermodynamic properties, including their size, shape, molar mass, degree of heterogeneity, oligomeric state, stoichiometry, and binding constants.

Device 

Proteomelab XL-I

Optical system 

absorbance + interference

Sample  

purified proteins and macromolecular complexes

Data/output

Determination of the sedimentation coefficient (S), the molecular weight, the dissociation constant (Kd), the stoechiometry of the complexe, etc

BIBLIOGRAPHY

Purification, characterization and crystallization of the human 80S ribosome. Khatter H, Myasnikov AG, Mastio L, Billas IM, Birck C, Stella S, Klaholz BP.Nucleic Acids Res. 2014 Apr;42(6):e49. doi: 10.1093/nar/gkt1404. Epub 2014 Jan 21.