Orientations of tyrosines 21 and 24 in coat subunits of Ff filamentous virus: determination by Raman linear intensity difference spectroscopy and implications for subunit packing.
Virions of the Ff group of bacteriophages (fd, f1, M13) are morphologically identical filaments (approximately 6-nm diameter x approximately 880-nm length) in which a covalently closed, single-stranded DNA genome is sheathed by approximately 2700 copies of a 50-residue alpha-helical subunit (pVIII). Orientations of pVIII tyrosines (Tyr21 and Tyr24) with respect to the filament axis have been determined by Raman linear intensity difference (RLID) spectroscopy of flow-oriented mutant virions in which the tyrosines were independently mutated to methionine. The results show that the twofold axis of the phenolic ring (C1-C4 line) of Tyr21 is inclined at 39.5 +/- 1.4 degrees from the virion axis, and that of Tyr24 is inclined at 43.7 +/- 0.6 degrees. The orientation determined for the Tyr21 phenol ring is close to that of a structural model previously proposed on the basis of fiber x-ray diffraction results (Protein Data Bank, identification code 1IFJ). On the other hand, the orientation determined for the Tyr24 phenol ring differs from the diffraction-based model by a 40 degrees rotation about the Calpha-Cbeta bond. The RLID results also indicate that each tyrosine mutation does not greatly affect the orientation of either the remaining tyrosine or single tryptophan (Trp26) of pVIII. On the basis of these results, a refined model is proposed for the coat protein structure in Ff.
Matsuno
M
M
Pharmaceutical Institute, Tohoku University, Sendai, Japan.
Takeuchi
H
H
Overman
S A
SA
Thomas
G J
GJ
D000595:Amino Acid Sequence; D017104:Bacteriophage M13 / Q000648:ultrastructure; D002213:Capsid / Q000737:chemistry*; D003090:Coliphages / Q000648:ultrastructure; D004277:DNA, Single-Stranded / Q000737:chemistry; D004279:DNA, Viral / Q000737:chemistry; D004926:Escherichia coli / Q000821:virology; D016679:Genome, Viral; D017912:Inovirus / Q000648:ultrastructure; D046911:Macromolecular Substances; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D013059:Spectrum Analysis, Raman; D014443:Tyrosine; D014771:Virion / Q000648:ultrastructure
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D046911:Macromolecular Substances; D014443:Tyrosine