Finding a needle in a haystack: detection of a small protein (the 12-kDa VP26) in a large complex (the 200-MDa capsid of herpes simplex virus).
Macromolecular complexes that consist of homopolymeric protein frameworks with additional proteins attached at strategic sites for a variety of structural and functional purposes are widespread in subcellular biology. One such complex is the capsid of herpes simplex virus type 1 whose basic framework consists of 960 copies of the viral protein, VP5 (149 kDa), arranged in an icosahedrally symmetric shell. This shell also contains major amounts of three other proteins, including VP26 (12 kDa), a small protein that is approximately equimolar with VP5 and accounts for approximately 6% of the capsid mass. With a view to inferring the role of VP26 in capsid assembly, we have localized it by quantitative difference imaging based on three-dimensional reconstructions calculated from cryo-electron micrographs. Purified capsids from which VP26 had been removed in vitro by treatment with guanidine hydrochloride were compared with preparations of the same depleted capsids to which purified VP26 had been rebound and with native (undepleted) capsids. The resulting three-dimensional density maps indicate that six VP26 subunits are distributed symmetrically around the outer tip of each hexon protrusion on VP26-containing capsids. Because VP26 may be readily dissociated from and reattached to the capsid, it does not appear to contribute significantly to structural stabilization. Rather, its exposed location suggests that VP26 may be involved in linking the capsid to the surrounding tegument and envelope at a later stage of viral assembly.
Booy
F P
FP
Laboratory of Structural Biology, National Institute of Arthritis, Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD 20892.
Trus
B L
BL
Newcomb
W W
WW
Brown
J C
JC
Conway
J F
JF
Steven
A C
AC
D002213:Capsid / Q000737:chemistry*; D036022:Capsid Proteins; D018259:Herpesvirus 1, Human / Q000737:chemistry / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008970:Molecular Weight
D036022:Capsid Proteins; D046911:Macromolecular Substances; C077409:capsid protein VP26, herpes simplex virus type 1; C018315:hexon capsid protein, Adenovirus; C043322:penton protein, adenovirus