We have identified a fifth member of the alpha subfamily of vertebrate laminin chains. Sequence analysis revealed a close relationship of alpha 5 to the only known Drosophila alpha chain, suggesting that the ancestral alpha gene was more similar to alpha 5 than to alpha 1-4. Analysis of RNA expression showed that alpha 5 is widely expressed in adult tissues, with highest levels in lung, heart, and kidney. Our results suggest that alpha 5 may be a major laminin chain of adult basal laminae.