Molecular characterization of the type-specific gamma-determinant located on the adenovirus fiber

Abstract

The fiber knob carries the type-specific gamma-antigen which can be demonstrated in hemagglutination inhibition tests. In order to characterize the gamma-determinant we selected subgenus DI adenovirus serotypes 9 and 19 (Ad9 and Ad19) which exhibited 29 amino acid exchanges in the knob domain. Like all subgenus DI adenoviruses they showed a complete hemagglutination pattern with rat and human erythrocytes. We constructed a total of 14 chimeric Ad9/Ad19 and Ad19/Ad9 fiber proteins, which possessed fiber knobs with progressively exchanged Ad9 and Ad19 amino acids. Furthermore, we created 39 fiber proteins with distinct amino acid exchanges in the knob regions by primer-directed mutagenesis. The proteins were expressed in Escherichia coli and tested in hemagglutination and hemagglutination inhibition tests. From our results we can conclude that the type-specific gamma-determinant is not restricted to a distinct region on the adenovirus fiber knob but is composed of at least 17 amino acids. Most of the amino acids contributing to the Ad9 and Ad19 gamma-determinants are located on the fiber knob loops.