Nanodisc Platform for Supported Membrane Protein Based Sensors and Catalysts

 

Incorporation of membrane proteins in functional biomaterials has been demonstrated for specific systems, but is difficult to consistently isolate and immobilize membrane proteins in their stabile and active state (1-4). We will develop a novel system to address this issue by first embedding the protein in small lipoprotein assemblies named nanodiscs, and then immobilizing the nanodiscs to silica particles via physisorption and covalent attachment with lipid-functional silanes (5,6). Diacylglycerol kinase (DGK) will be used as the model membrane protein during the development of the system (7). For Theme 1, the REU student will learn basic biochemical and molecular biology techniques to assemble the nanodisc and conduct activity assay in Dr. Wei’s lab. For Theme 2, the student will investigate the immobilization of the protein-incorporated nanodiscs to micron-scale mesoporous silica particles and characterize the location and activity of the immobilized protein in the labs of Dr. Rankin and Knutson.


 

1. Wagner ML, Tamm LK (2000) Tethered polymer-supported planar lipid bilayers for reconstitution of integral membrane proteins: silane-polyethyleneglycol-lipid as a cushion and covalent linker. Biophys J 79: 1400–1414.

2. Sackmann E (1996) Supported membranes: scientific and practical applications. Science 271: 43–48.

3. Richter RP, Berat R, Brisson AR (2006) Formation of solid-supported lipid bilayers: an integrated view. Langmuir 22: 3497–3505.

4. Kusters I, Mukherjee N, de Jong MR, Tans S, Koçer A, et al. (2011) Taming Membranes: Functional Immobilization of Biological Membranes in Hydrogels. PLoS ONE 6(5): e20435. doi:10.1371/journal.pone.0020435

5. Schuler, M. A., Denisov, I. G., Sligar, S. G. (2013) “Nanodiscs as a new tool to examine lipid-protein interactions.” Methods Mol Biol. 974, 415-433.

6. Sloan, C. D., Marty, M. T., Sligar, S. G., Bailey, R. C. (2013) “Interfacing lipid bilayer nanodiscs and silicon photonic sensor arrays for multiplexed protein-lipid and protein-membrane protein interaction screening.” Anal. Chem., 85(5), 2970-2976.

7. Li, DF, Lyons, JA, Pye, VE, Vogeley, L, Aragao, D, Kenyon, CP, Shah, STA, Doherty, C, Aherne, M, Caffrey, M, Crystal structure of the integral membrane diacylglycerol kinase, Nature, 497, 7450, 2013, 521-524