The maturation-dependent conformational change of phage T4 capsid involves the translocation of specific epitopes between the inner and the outer capsid surfaces.
After polymerization of the phage T4 prohead is complete, its capsid expands by approximately 16%, is greatly stabilized, and acquires the capacity to bind accessory proteins. These effects are manifestations of a large-scale, irreversible, conformational change undergone by the major capsid protein, gp23 (521 residues) which is cleaved to gp23* (residues 66-521) during this maturation process. In order to explore its structural basis, we have performed immunoelectron microscopy with antibodies raised against synthetic peptides that correspond to precisely defined segments of the amino acid sequence of gp23. These antibodies were used to label purified polyheads (tubular polymorphic variants of the normal icosahedral capsid), in experiments designed to impose constraints on the possible foldings of the gp23/gp23* polypeptide chains in their successive conformational states. Peptide 1 (residues 48-57), part of the gp23-delta domain that is excised when gp23 is converted to gp23*, resides on the inner surface of the precursor surface lattice, but--if not proteolyzed--is found on the outer surface of the mature surface lattice. Peptide 2 (residues 65-73), immediately distal to the cleavage site, is located on the inside of the precursor surface lattice, and remains there subsequent to expansion. Peptide 3 (residues 139-146) is translocated in the opposite direction from peptide 1, i.e., from the outer to the inner surface upon expansion; moreover, expansion greatly increases the polyheads' affinity for these antibodies. Peptide 5 (residues 301-308) is located on the inside in both the precursor and the mature states. Taking into account data from other sources, these observations imply that the conformational change that underlies capsid expansion involves a radical reorganization of the proteins' structure, in which at least three distinct epitopes, situated in widely differing parts of the polypeptide chain, are translocated from one side to the other. Moreover, the amino-terminal portion of gp23/gp23*, around the cleavage site, is particularly affected.
Steven
A C
AC
Laboratory of Structural Biology Research, National Institute of Arthritis, Musculoskeletal, and Skin Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Bauer
A C
AC
Bisher
M E
ME
Robey
F A
FA
Black
L W
LW
D000595:Amino Acid Sequence; D000906:Antibodies / Q000302:isolation & purification; D002213:Capsid / Q000276:immunology / Q000648:ultrastructure*; D036022:Capsid Proteins; D000939:Epitopes; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011498:Protein Precursors; D013329:Structure-Activity Relationship; D013604:T-Phages / Q000648:ultrastructure*; D014764:Viral Proteins / Q000276:immunology / Q000648:ultrastructure*; D014779:Virus Replication / Q000502:physiology*
D000906:Antibodies; D036022:Capsid Proteins; D000939:Epitopes; D011498:Protein Precursors; D014764:Viral Proteins; C064517:gp23 protein, Bacteriophage T4