Abstract

The membrane bound forms of the coat proteins of the filamentous bacteriophages fd and Pfl will be studied by high resolution NMR spectroscopy. The proposed research is highly focused on obtaining structural and dynamical information about these proteins from one- and two-dimensional 1H and 15N NMR experiments. Preliminary 1H two-dimensional correlated and NOE spectra show many cross-peaks, including those between N-1H amd CAlpha - 1H resonances, indicating tht proton connectivities can be mapped out on both through-bond and through-space bases. The secondary structure of the immobile protein segments will be determined from these 1H NMR experiments. Well resolved 15N spectra enable the relaxation parameters, especially the heteronuclear NOE, to be used to describe protein dynamics. The availability of specifically 15N labelled proteins, the use of heteronuclear two-dimensional correlations, and the generation of site specific mutations will assist in making both 1H and 15N resonance assignments. The properties of the coat proteins in membrane environments will be compared to those in the assembled virus.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI020770-03
Application #
3130580
Study Section
Biophysics and Biophysical Chemistry B Study Section (BBCB)
Project Start
1984-12-01
Project End
1987-11-30
Budget Start
1986-12-01
Budget End
1987-11-30
Support Year
3
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
University of Pennsylvania
Department
Type
Schools of Arts and Sciences
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104

  Publications

Kim, Y; Valentine, K; Opella, S J et al. (1998) Solid-state NMR studies of the membrane-bound closed state of the colicin E1 channel domain in lipid bilayers. Protein Sci 7:342-8
Marassi, F M; Ramamoorthy, A; Opella, S J (1997) Complete resolution of the solid-state NMR spectrum of a uniformly 15N-labeled membrane protein in phospholipid bilayers. Proc Natl Acad Sci U S A 94:8551-6
Almeida, F C; Opella, S J (1997) fd coat protein structure in membrane environments: structural dynamics of the loop between the hydrophobic trans-membrane helix and the amphipathic in-plane helix. J Mol Biol 270:481-95
Gesell, J; Zasloff, M; Opella, S J (1997) Two-dimensional 1H NMR experiments show that the 23-residue magainin antibiotic peptide is an alpha-helix in dodecylphosphocholine micelles, sodium dodecylsulfate micelles, and trifluoroethanol/water solution. J Biomol NMR 9:127-35
Almeida, F C; Opella, S J (1997) Measurement of 1H T1 rho in a uniformly 15N-labeled protein in solution with heteronuclear two-dimensional spectroscopy. J Magn Reson 124:509-11
Klassen, R B; Opella, S J (1997) NMR studies of peptides and proteins associated with membranes. Methods Mol Biol 60:271-97
Howard, K P; Opella, S J (1996) High-resolution solid-state NMR spectra of integral membrane proteins reconstituted into magnetically oriented phospholipid bilayers. J Magn Reson B 112:91-4
Bechinger, B; Gierasch, L M; Montal, M et al. (1996) Orientations of helical peptides in membrane bilayers by solid state NMR spectroscopy. Solid State Nucl Magn Reson 7:185-91
Tobias, D J; Gesell, J; Klein, M L et al. (1995) A simple protocol for identification of helical and mobile residues in membrane proteins. J Mol Biol 253:391-5
Opella, S J; Kim, Y; McDonnell, P (1994) Experimental nuclear magnetic resonance studies of membrane proteins. Methods Enzymol 239:536-60

Showing the most recent 10 out of 19 publications