Purification in quantity of the secreted form of WI-1: a major adhesin on Blastomyces dermatitidis yeasts.

WI-1, a 120-kDa adhesin on Blastomyces dermatitidis, binds the yeast to macrophages and is a major target antigen of immune recognition in acquired resistance to the fungus. In past studies, WI-1 has been purified by extracting the protein from the yeast cell wall, which yields microgram quantities for biological assays. We report a strategy for generating and purifying the secreted form of WI-1 in quantity. Yeasts of B. dermatitidis ATCC strain 60636 cultured in HMM medium were found to secrete 10 microg/ml of WI-1 into a supernate relatively free of other medium and yeast components. Using a two-step method of ion exchange and hydrophobic interaction chromatography, we achieved a 7.1-fold purification of WI-1. Purified WI-1 was sequenced at the N-terminus which revealed that the secreted protein exists in two different forms. In functional assays, purified WI-1 also retained its adhesivity for human macrophages, and its antigenicity in binding anti-WI-1 antibodies and stimulating T-cells to proliferate, but it lost some capacity to elicit delayed-type hypersensitivity in mice. These findings advance our understanding of the WI-1 adhesin/antigen and our ability to express and purify WI-1 in quantity and will permit a study of the relationship between three-dimensional structure and activity of the molecule. Copyright 1997 Academic Press.