A robust method to screen detergents for membrane protein stabilization, revisited.

Philippe Champeil (1, 2) , Stéphane Orlowski (1, 2) , Simon Babin (1, 2) , Sten Lund (3) , Marc Le Maire (1, 2) , Jesper Møller (4, 5) , Guillaume Lenoir (1, 2) , Cédric Montigny (1, 2)

1 LPSM - Laboratoire des Protéines et Systèmes Membranaires
2 IBITECS - Institut de Biologie et de Technologies de Saclay
3 Medical Research Laboratory, Department of Endocrinology and Internal Medicine
4 PUMPKIN - Centre for Membrane Pumps in Cells and Disease
5 Department of Biomedicine

This report is a follow up of our previous paper (Lund, Orlowski, de Foresta, Champeil, le Maire and Møller (1989), J Biol Chem 264:4907–4915) showing that solubilization in detergent of a membrane protein may interfere with its long-term stability, and proposing a protocol to reveal the kinetics of such irreversible inactivation. We here clarify the fact that when various detergents are tested for their effects, special attention has of course to be paid to their critical micelle concentration. We also investigate the effects of a few more detergents, some of which have been recently advertised in the literature, and emphasize the role of lipids together with detergents. Among these detergents, lauryl maltose neopentyl glycol (LMNG) exerts a remarkable ability, even higher than that of β-dodecylmaltoside (DDM), to protect our test enzyme, the paradigmatic P-type ATPase SERCA1a from sarcoplasmic reticulum. Performing such experiments for one's favourite protein probably remains useful in pre-screening assays testing various detergents.

Mots clés: Detergent; Membrane protein; Lipid; Stability; LMNG; SERCA1a

Domaines: Sciences du Vivant [q-bio]

Philippe Champeil, Stéphane Orlowski, Simon Babin, Sten Lund, Marc Le Maire, et al.. A robust method to screen detergents for membrane protein stabilization, revisited.. Analytical Biochemistry, 2016, 511, pp.31-35. ⟨hal-01356014⟩