Bead-linked proteoliposomes: a reconstitution method for nmr analyses of membrane protein-ligand interactions.
Structural information about the interactions between membrane proteins and their ligands provides insights into the membrane protein functions. A variety of surfactants have been used for structural analyses of membrane proteins, and in some cases, they yielded successful results. However, the use of surfactants frequently increases the conformational instability of membrane proteins and distorts their normal function. Here, we propose a new strategy of membrane protein reconstitution into lipid bilayers on affinity beads, which maintains the native conformation and function of the protein for NMR studies. The reconstituted membrane proteins are suitable for NMR analyses of interactions, by using the transferred cross-saturation method. The strategy was successfully applied to the interaction between a potassium ion channel, KcsA, and a pore-blocker, agitoxin2 (AgTx2). This strategy would be useful for analyzing the interactions between various membrane proteins and their ligands.
Yokogawa
Mariko
M
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Takeuchi
Koh
K
Shimada
Ichio
I
D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010759:Phosphorus Isotopes; D015221:Potassium Channels / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011510:Proteolipids / Q000737:chemistry* / Q000378:metabolism; D012604:Scorpion Venoms / Q000737:chemistry / Q000378:metabolism; D013501:Surface-Active Agents / Q000737:chemistry*
D001426:Bacterial Proteins; D008024:Ligands; D008565:Membrane Proteins; D010759:Phosphorus Isotopes; D015221:Potassium Channels; D011510:Proteolipids; D012604:Scorpion Venoms; D013501:Surface-Active Agents; C107790:prokaryotic potassium channel; C020485:proteoliposomes; C087603:agitoxin 2