Assignment of 1H, 15N, and backbone 13C resonances in detergent-solubilized M13 coat protein via multinuclear multidimensional NMR: a model for the coat protein monomer.
The major coat protein (gVIIIp) of bacteriophage M13 complexed with SDS detergent micelles was used as a model system to study the lipid-bound conformation of the protein. Conditions were found that allowed the recording of good quality of NMR spectra. By making extensive use of three-dimensional heteronuclear (13C, 15N) NMR, we obtained a complete set of resonance assignments for 1HN, 1H alpha, 1H beta, 13C alpha, CO, and 15N and partially assigned the rest of the 1H spectrum. Analysis of NOE and chemical shift data reveals that gVIIIp is composed of two alpha-helical domains, one ranging from Pro-6 to Glu20 and the other ranging from Tyr-24 all the way to the C-terminus Ser-50. In contrast to the results reported by Henry and Sykes [Henry, G.D., & Sykes, B.D. (1992) Biochemistry 31, 5285-5297], at a high SDS to protein ratio the protein appears to be monomeric.
van de Ven
F J
FJ
Laboratory of Biophysical Chemistry, NSR Center, University of Nijmegen, The Netherlands.
van Os
J W
JW
Aelen
J M
JM
Wymenga
S S
SS
Remerowski
M L
ML
Konings
R N
RN
Hilbers
C W
CW
D000595:Amino Acid Sequence; D017104:Bacteriophage M13 / Q000737:chemistry / Q000235:genetics; D002213:Capsid / Q000737:chemistry* / Q000378:metabolism; D036022:Capsid Proteins; D004926:Escherichia coli / Q000235:genetics; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D008563:Membrane Lipids / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D008823:Micelles; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate; D012995:Solubility
D036022:Capsid Proteins; D046911:Macromolecular Substances; D008563:Membrane Lipids; D008565:Membrane Proteins; D008823:Micelles; D011994:Recombinant Proteins; C025668:coat protein, Bacteriophage M13; D012967:Sodium Dodecyl Sulfate