Bicelles in structure-function studies of membrane-associated proteins.
Bicelles are a novel form of long-chain/short-chain phospholipid aggregates, which are useful for biophysical and biochemical studies of membrane-associated biomolecules. In this work, we review the development of bicelles and their uses in structural characterization (primarily via NMR, circular dichroism, and fluorescence) of membrane-associated peptides. We also show that bicellar phospholipids are substrates for lipolytic enzymes. For this latter work, we employed a 31P NMR enzymatic assay system to examine the kinetic behavior of cobra venom phospholipase A(2) toward a variety of bicellar substrates. This enzyme hydrolyzed all bicelle lipids at rates comparable to those found for the enzyme action on traditional micellar substrates, which are the best substrates for this enzyme. In addition, we found that this PLA(2) showed no significant preference for long-chain or short-chain phospholipids when they were presented as mixtures in bicelles.
Whiles
Jennifer A
JA
Department of Chemistry and Biochemistry, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0601, USA.
Deems
Raymond
R
Vold
Regitze R
RR
Dennis
Edward A
EA
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000378:metabolism; D000818:Animals; D002942:Circular Dichroism; D004134:Dimyristoylphosphatidylcholine / Q000378:metabolism; D004546:Elapid Venoms / Q000737:chemistry; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008563:Membrane Lipids / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D008968:Molecular Conformation; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D010741:Phospholipases A / Q000737:chemistry* / Q000378:metabolism*; D010743:Phospholipids / Q000737:chemistry*; D010843:Pichia; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D014162:Transfection
D004546:Elapid Venoms; D008563:Membrane Lipids; D008565:Membrane Proteins; D010455:Peptides; D010743:Phospholipids; D011994:Recombinant Proteins; D015060:1,2-Dipalmitoylphosphatidylcholine; D010741:Phospholipases A; D004134:Dimyristoylphosphatidylcholine