High resolution two-dimensional gel electrophoresis of structural proteins of baculoviruses of Autographa californica and Porthetria (lymantria) dispar.

The structural polypeptides of baculoviruses of Autographa californica (AcMNPV) and Porthetria dispar (PdMNPV) were analyzed by two-dimensional (2-D) gel electrophoresis. Purified proteins were solubilized in urea-NP40 mix and separated by isoelectric focusing in the first dimension; electrophoresis in the presence of sodium dodecyl sulfate (SDS) separated proteins by molecular weight in the second dimension. Eighty-one acidic polypeptides ranging in molecular weight from 13,500 to 86,000 Da were resolved in AcMNPV enveloped virions. The predominant polypeptide had a molecular weight of 41,500 and was considered to be the major capsid protein. Nucleocapsids from AcMNPV were resolved into 64 polypeptides. At least 11 of the polypeptides, including most of the high molecular weight proteins, that were not resolved in nucleocapsids were considered to be envelope proteins. For PdMNPV enveloped virions, there were 95 acidic polypeptides ranging in molecular weight from 13,500 to 85,500. The predominant polypeptide had a molecular weight of 46,500 Da. Polyhedral proteins (polyhedrin) isolated from protease-inactivated polyhedra and separable into a single major polypeptide (approx. 31,000) on one-dimensional SDS-polyacrylamide gel electrophoresis were resolved into six polypeptides for both viruses. All six polyhedrin polypeptides had the same molecular weight, but their isoelectric points ranged from pH 5.3 to 5.9 for AcMNPV and from pH 5.7 to 6.2 for PdMNPV. These six polypeptides were also detected when protease-inactivated or noninactivated whole polyhedra were analyzed directly by 2-D electrophoresis. It is assumed that not all the observed baculovirus polypeptides were unique species. Some proteins, especially the polyhedrin polypeptides, appeared to be related and had altered mobilities as a consequence of post-translational modifications.