|David P Hoogerheide
Localization of proteins to bilayer lipid membranes (BLMs) is a complex but fundamental cellular process. The mechanism of binding varies widely among proteins and is difficult to elucidate due to the many actors involved in the binding process. The bioreflectometry group at the NIST Center for Neutron Research collaborates with academic and various government laboratories to elucidate mechanisms of protein binding to BLMs. We comprise researchers with a broad range of expertise and are actively developing advanced biochemical and biophysical techniques, particularly those useful for neutron scattering, to study membrane-associated proteins. We are interested in chemistries for new measurement platforms, protein synthesis and diagnostics, advanced data analysis techniques, instrument and sample environment development, and simulation methods to compare to experimental results.
We are particularly interested in the development of two techniques based on a planar lipid bilayer architecture to complement neutron scattering studies on peripheral membrane protein structure. The first, bilayer overtone analysis, involves measurement of the electric field-induced compression of a BLM to detect asymmetries in a membrane caused by binding of analyte molecules. The second, reconstitution of biological nanopores into the BLM, allows the single-molecule probing of membrane binding properties, including binding energy. Both techniques have been successfully applied to understanding the mechanisms of membrane binding of the cytosolic mitochondrial regulators α-synuclein and dimeric tubulin, and we are poised to apply them to other peripheral membrane protein systems.
Hoogerheide DP, Noskov SY, Jacobs D, Bergdoll L, Silin V, Worcester DL, Abramson J, Nanda H, Rostovtseva TK, Bezrukov SM: “Structural features and lipid binding domain of tubulin on biomimetic mitochondrial membranes.” Proceedings of the National Academy of Sciences of the USA 114(18): E3622-E3631, 2017
Hoogerheide DP, Gurnev PA, Rostovtseva TK, Bezrukov SM: “Mechanism of α-synuclein translocation through a VDAC nanopore revealed by energy landscape modeling of escape time distributions.” Nanoscale 9: 183-192, 2017
Rostovtseva TK, Hoogerheide DP, Rovini A, Bezrukov SM: “Lipids in Regulation of the Mitochondrial Outer Membrane Permeability, Bioenergetics, and Metabolism.” in Molecular Basis for Mitochondrial Signaling (Biological and Medical Physics, Biomedical Engineering). Rostovtseva TK, editor. Springer International (Cham, Switzerland), 2017
Binding (biochemical); Lipid bilayers; Membrane proteins; Nanopores; Bilayer overtone analysis; Biophysical techniques; Single molecule techniques; Planar bilayer lipid membranes;