Genes for the MSP1a and MSP1b subunits of the Anaplasma marginale surface antigen complex MSP1 were previously cloned and expressed in Escherichia coli. We report here the localization of MSP1a and MSP1b polypeptides on the surface of recombinant E. coli by using a live cell indirect immunofluorescent antibody assay. Recombinant E. coli cells expressing the msp1 alpha gene or the msp1 beta gene encoding the MSP1a and MSP1b polypeptide subunits, respectively, were shown by a culture recovery adhesion assay and by direct microscopic examination to specifically adhere to bovine erythrocytes. This adhesion was more than additive when both genes were coexpressed in a single recombinant construct. Similarly, these recombinants hemagglutinated bovine erythrocytes in a microtiter hemagglutination assay. Inhibition of recombinant E. coli adhesion to bovine erythrocytes and hemagglutination inhibition were observed in the presence of homologous monospecific polyclonal antiserum raised against purified MSP1a or MSP1b polypeptide. These data suggest that the MSP1a and MSP1b polypeptides have functions as adhesins on A. marginale initial bodies, probably during erythrocyte invasion.