Berget, PB and King, J. (1978) J Mol Biol,. 124(3), 469-486.
Isolation and characterization of precursors in T4 baseplate assembly: The complex of gene 10 and gene 11 products.
The first multi-protein precursor in the assembly of the radial arms of the T4 baseplate has been purified to homogeneity. The complex was isolated from cells infected with a mutant blocked in the subsequent step in baseplate arm assembly. The assay for this precursor exploited in the fact that the complex contains the target antigen of the neutralizing antibodies found in antibaseplate serum (Berget & King, 1978).
The complex is composed of gene 10 protein (Mr, 88,000) and gene 11 protein (Mr, 24,000). Analytical ultracentrifugation experiments revealed a molecular weight of 258,000 and a sedimentation coefficient of 9.3 S for the complex. The overall and single polypeptide chain molecular weights are consistent with the complex containing two gene 10 polypeptides. Visualization of the ccomplex in the electron microscope revealed an asymmetric angular structure. The shape, together with the previous identification of gene 11 product as the tailspike protein (Crowther te al., 1977), indicates that the complex forms the body of the spikes and vertices of the hexagonal baseplate.
Using an in vitro baseplate assembly assay, it was possible to demonstrate that the complex contains both the assembly-active gene 10 and gene 11 products. Gene 11 product (from 10- extracts) can convert 11- particles to viable phage. However, the complex lacked this activity, indicating that it does not readily dissociate. The preccursor complex could be dissociated with denaturing solvents. Upon returning to physiological conditions, both the antigenic and biological activities of the gene 11 product could be recovered. The biological activity of the gene 10 product was not regained.