Protein and small molecule recognition properties of deep cavitands in a supported lipid membrane determined by calcination-enhanced SPR spectroscopy.
This paper details the incorporation of a water-soluble deep cavitand into a membrane bilayer assembled onto a nanoglassified surface for study of molecular recognition in a membrane-mimicking setting. The cavitand retains its host properties, and real-time analysis of the host:guest properties of the membrane:cavitand complex via surface plasmon resonance and fluorescence microscopy is described. The host shows selectivity for choline-derived substrates, and no competitive incorporation of substrate is observed in the membrane bilayer. A variety of trimethylammonium-derived substrates are suitable guests, displaying varied binding affinities in a millimolar range. The membrane:cavitand:guest complexes can be subsequently used to capture NeutrAvidin protein at the membrane surface if a biotin-derived guest molecule is used. The surface coverage of NeutrAvidin is affected by the spacer used to derivatize the biotin. Increased distance from the bilayer allows a higher concentration of protein to be immobilized, suggesting a diminishing detrimental steric effect when the binding event is shifted away from the surface.
Liu
Ying
Y
Department of Chemistry, University of California, Riverside, California 92521, USA.
Liao
Puhong
P
Cheng
Quan
Q
Hooley
Richard J
RJ
D000818:Animals; D001360:Avidin / Q000378:metabolism*; D002118:Calcium / Q000737:chemistry; D002794:Choline / Q000031:analogs & derivatives / Q000378:metabolism*; D004988:Ethers, Cyclic / Q000737:chemistry / Q000378:metabolism*; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D011485:Protein Binding; D012118:Resorcinols / Q000737:chemistry / Q000378:metabolism*; D020349:Surface Plasmon Resonance / Q000379:methods*; D013499:Surface Properties
D004988:Ethers, Cyclic; D008051:Lipid Bilayers; D012118:Resorcinols; C442939:cavitand; C093483:neutravidin; D001360:Avidin; D002794:Choline; D002118:Calcium