Amphipols can support the activity of a membrane enzyme.
Amphipathic polymers ("amphipols") were introduced several years ago (Tribet, C.; Audebert, R.; Popot, J.-L. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 15047-15050) as an alternative method for solubilizing integral membrane proteins in stable, nativelike conformations. However, direct maintenance of full membrane protein functionality in amphipol solutions has not previously been demonstrated in the absence of added lipid or detergent. In this contribution, the first zwitterionic amphipol "PMAL-B-100" is introduced. PMAL-B-100 not only maintains membrane protein structure and solubility, but also supports the full catalytic activity of an integral membrane enzyme, diacylglycerol kinase, in the complete absence of additional lipid or detergent. All of the roles which a lipid bilayer normally plays in maintaining diacylglycerol kinase's structure and in facilitating catalysis are satisfied by the environment and interactions supplied by PMAL-B-100.
Gorzelle
Bonnie M
BM
Department of Physiology and Biophysics and Cleveland Center for Structural Biology, Case Western Reserve University, Cleveland, OH 44106-4970, USA.
Hoffman
Amy Kuhn
AK
Keyes
Melvin H
MH
Gray
Don N
DN
Ray
Dale G
DG
Sanders
Charles R
CR
D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D002384:Catalysis; D019852:Diacylglycerol Kinase / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D007700:Kinetics; D008081:Liposomes; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008823:Micelles; D011108:Polymers / Q000737:chemistry* / Q000494:pharmacology
D008081:Liposomes; D008565:Membrane Proteins; D008823:Micelles; D011108:Polymers; D000255:Adenosine Triphosphate; D019852:Diacylglycerol Kinase