Protocol

Membrane Biogenesis

Volume 1033 of the series Methods in Molecular Biology pp 219-251

Date:

Sedimentation Velocity Analytical Ultracentrifugation in Hydrogenated and Deuterated Solvents for the Characterization of Membrane Proteins

  • Aline Le RoyAffiliated withInstitut de Biologie Structurale, CEA
  • , Hugues NuryAffiliated withInstitut de Biologie Structurale, CEA
  • , Benjamin WisemanAffiliated withInstitut de Biologie Structurale, CEA
  • , Jonathan SarwanAffiliated withInstitut de Biologie Structurale, CEA
  • , Jean-Michel JaultAffiliated withInstitut de Biologie Structurale, CEA
  • , Christine EbelAffiliated withInstitut de Biologie Structurale, CEA

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Abstract

This chapter is a step-by-step protocol for setting up, realizing, and analyzing sedimentation velocity experiments in hydrogenated and deuterated solvents, in the context of the characterization of membrane protein, in terms of homogeneity, association state, and amount of bound detergent, based on a real case study of the membrane protein BmrA solubilized in n-Dodecyl-β-d-Maltopyranoside) detergent.

Key words

Sedimentation velocity Analytical ultracentrifugation Membrane proteins Homogeneity Association state Bound detergent BmrA Detergent Heavy water D2O SEDFIT