Presented here is a new approach for the elucidation of the dynamics and structure of membrane bound proteins and polypeptides with atomic resolution that is based on solid state Nuclear Magnetic Resonance experiments. The structural aspects of the effort result from determining the orientation of covalent bonds within the polymerws with respect to a common axis, the magnetic field direction. From the bond orientations of adjacent groups the torsion angles are determined defining the relative orientation of these groups. The dynamic characterization is composed of two parts; first a spatial characterization of the motion, defining the axis about which the motion occurs, the type of motion whether it is diffusional or discontinuous as in flip motions between two potential minima, and finally the amplitude of the motion. Only when this spatial model for the motion is complete can an accurate determination of the frequency of the motion be made from the NMR data. To demonstrate the validity of this approach for determining structure and dynamics and to answer many biophysical questions concerning the function of the Gramicidin A cation channel at an atomic level this molecule will be studied. The ion transport characteristics of the channel have been extensively studied in the several laboratories. These results show that the Gramicidini channel may be a very useful model for the proteinaceous Na+ and K+ channels of higher organisms. The folding motiff for the polypeptide backbone is generally accepted as that of the original channel model. However, the structural details are unresolved at this time and the magnitude of the structural distortions that take place upon binding ions is not known. Structural studies have been hindered both by the extreme sensitivity if this molecule to its solvent environment and by the difficulty of forming cocfrystals of Gramicfidin and lipid which will diffract to high resolution. Furthermore, the dynamics of the atoms in the polypeptide backbone which presumably coordinate the unhydrated ions during passage through the channel are uncharacterized. With both a detailed structural and dynamic picture of Gramicidin unprecedented correlations between structure, dynamics and function will be achieved.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI023007-01A1
Application #
3134816
Study Section
Biophysics and Biophysical Chemistry B Study Section (BBCB)
Project Start
1987-01-01
Project End
1989-12-31
Budget Start
1987-01-01
Budget End
1987-12-31
Support Year
1
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Florida State University
Department
Type
Schools of Arts and Sciences
DUNS #
020520466
City
Tallahassee
State
FL
Country
United States
Zip Code
32306
Chipot, Christophe; Dehez, François; Schnell, Jason R et al. (2018) Perturbations of Native Membrane Protein Structure in Alkyl Phosphocholine Detergents: A Critical Assessment of NMR and Biophysical Studies. Chem Rev 118:3559-3607
Qin, Huajun; Miao, Yimin; Cross, Timothy A et al. (2017) Beyond Structural Biology to Functional Biology: Solid-State NMR Experiments and Strategies for Understanding the M2 Proton Channel Conductance. J Phys Chem B 121:4799-4809
Fu, Riqiang; Miao, Yimin; Qin, Huajun et al. (2016) Probing Hydronium Ion Histidine NH Exchange Rate Constants in the M2 Channel via Indirect Observation of Dipolar-Dephased 15N Signals in Magic-Angle-Spinning NMR. J Am Chem Soc 138:15801-15804
Wright, Anna K; Batsomboon, Paratchata; Dai, Jian et al. (2016) Differential Binding of Rimantadine Enantiomers to Influenza A M2 Proton Channel. J Am Chem Soc 138:1506-9
Ekanayake, E Vindana; Fu, Riqiang; Cross, Timothy A (2016) Structural Influences: Cholesterol, Drug, and Proton Binding to Full-Length Influenza A M2 Protein. Biophys J 110:1391-9
Gleed, Mitchell L; Ioannidis, Harris; Kolocouris, Antonios et al. (2015) Resistance-Mutation (N31) Effects on Drug Orientation and Channel Hydration in Amantadine-Bound Influenza A M2. J Phys Chem B 119:11548-59
Gleed, Mitchell L; Busath, David D (2015) Why bound amantadine fails to inhibit proton conductance according to simulations of the drug-resistant influenza A M2 (S31N). J Phys Chem B 119:1225-31
Miao, Yimin; Fu, Riqiang; Zhou, Huan-Xiang et al. (2015) Dynamic Short Hydrogen Bonds in Histidine Tetrad of Full-Length M2 Proton Channel Reveal Tetrameric Structural Heterogeneity and Functional Mechanism. Structure 23:2300-2308
Das, Nabanita; Dai, Jian; Hung, Ivan et al. (2015) Structure of CrgA, a cell division structural and regulatory protein from Mycobacterium tuberculosis, in lipid bilayers. Proc Natl Acad Sci U S A 112:E119-26
Cross, Timothy A; Ekanayake, Vindana; Paulino, Joana et al. (2014) Solid state NMR: The essential technology for helical membrane protein structural characterization. J Magn Reson 239:100-9

Showing the most recent 10 out of 99 publications