3.3. Background and Significance 3.3.1. There is a compelling need for novel model membrane media that are optimized for MP structural biology. The vast majority of MP structures determined to date by either NMR or crystallographic methods were determined using classical detergent as the model membrane media. Unfortunately, even mild detergents destabilize MPs relative to their stability in native membranes. Moreover, many of the mildest detergents (such as the alkyl maltoside) routinely fail to yield high quality NMR spectra. More native-like media, such as detergent-lipid mixed micelles or bicelles sometimes offer advantages, but also have drawbacks. Moreover, while membranes from higher organisms almost always contain a considerable mole fraction of cholesterol, very little work has been carried out to establish NMR-compatible media that contain cholesterol, in part because of its very low solubility in detergent solutions. It is clear that there is a need to expand the range of membrane-mimetic media that are available. We propose to design, synthesize (through Core B) and test new surfactants for use in structural biological studies of MPs. We also will test surfactants and cholesterol analogs that have recently been commercialized but not yet well tested.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Specialized Center--Cooperative Agreements (U54)
Project #
5U54GM094608-05
Application #
8688796
Study Section
Special Emphasis Panel (ZGM1-CBB-3)
Project Start
Project End
Budget Start
2014-07-01
Budget End
2015-06-30
Support Year
5
Fiscal Year
2014
Total Cost
$378,331
Indirect Cost
$86,408
Name
Harvard Medical School
Department
Type
DUNS #
047006379
City
Boston
State
MA
Country
United States
Zip Code
02115
Kim, Ji-Hun; Schlebach, Jonathan P; Lu, Zhenwei et al. (2016) A pH-Mediated Topological Switch within the N-Terminal Domain of Human Caveolin-3. Biophys J 110:2475-2485
Schlebach, Jonathan P; Barrett, Paul J; Day, Charles A et al. (2016) Topologically Diverse Human Membrane Proteins Partition to Liquid-Disordered Domains in Phase-Separated Lipid Vesicles. Biochemistry 55:985-8
Oxenoid, Kirill; Dong, Ying; Cao, Chan et al. (2016) Architecture of the mitochondrial calcium uniporter. Nature 533:269-73
Hagn, Franz; Wagner, Gerhard (2015) Structure refinement and membrane positioning of selectively labeled OmpX in phospholipid nanodiscs. J Biomol NMR 61:249-60
Schlebach, Jonathan P; Sanders, Charles R (2015) Influence of Pathogenic Mutations on the Energetics of Translocon-Mediated Bilayer Integration of Transmembrane Helices. J Membr Biol 248:371-81
Williamson, Jessica A; Cho, Seung-Hyun; Ye, Jiqing et al. (2015) Structure and multistate function of the transmembrane electron transporter CcdA. Nat Struct Mol Biol 22:809-14
Schlebach, Jonathan P; Sanders, Charles R (2015) The safety dance: biophysics of membrane protein folding and misfolding in a cellular context. Q Rev Biophys 48:1-34
Dev, Jyoti; Br├╝schweiler, Sven; Ouyang, Bo et al. (2015) Transverse relaxation dispersion of the p7 membrane channel from hepatitis C virus reveals conformational breathing. J Biomol NMR 61:369-78
Chatterjee, Deep; Eckert, Carl Elias; Slavov, Chavdar et al. (2015) Influence of Arrestin on the Photodecay of Bovine Rhodopsin. Angew Chem Int Ed Engl 54:13555-60
Br├╝schweiler, Sven; Yang, Qin; Run, Changqing et al. (2015) Substrate-modulated ADP/ATP-transporter dynamics revealed by NMR relaxation dispersion. Nat Struct Mol Biol 22:636-41

Showing the most recent 10 out of 61 publications