Serological homologies between H1 degrees and H5 include the carboxyl-terminal domain

We have reported previously that antibodies to chicken H5 and antibodies to H1 both cross-react with mammalian H1 degree (Mura, C. V., and Stollar, B. D. (1981) J. Biol. Chem. 256, 9767-9769). The antigenic sites in H1 degree recognized by these antibodies were analyzed using immunoblotting. Peptides of H1 degree were prepared by partial digestion with acetic acid and tested for reactivity with: 1) antibodies induced by H5 alone, which reacted primarily with the central globular region of H5; 2) antibodies induced by H5 X RNA complexes, which reacted with this domain as well as the basic COOH-terminal domain; and 3) antiserum to calf thymus H1. Anti-H5 antibodies (anti-globular region) cross-reacted with H1 degree peptides that co-migrated with peptides of H5 that contain the globular region, but did not cross-react with H1. Anti-H5/RNA antibodies (anti-globular + anti-COOH-terminal) cross-reacted with these peptides and, in addition, with a lysine-rich H1 degree peptide that co-migrated with the basic COOH-terminal H5 peptide. This H1 degree peptide, but not the putative globular H1 degree peptides, was also recognized by an antiserum to calf H1 which was primarily reactive with the large, COOH-terminal N-bromosuccinimide fragment of calf H1. A weaker cross-reaction between this antiserum and the carboxyl-terminal domain of H5 could be visualized when large quantities of H5 were used in immunoblots. The results indicate that structural homologies between H5 and H1 degree extend beyond the globular region and into the lysine-rich carboxyl-terminal domain. Antigenic homologies between H1 degree and H1 are also at least partially localized in this domain. H1 degree is serologically intermediate between H5 and H1.