Alkaline cell extracts obtained from whole cells of a flocculent strain of Saccharomyces cerevisiae, containing the dominant gene for flocculence FL04, and a nonflocculent mutant (FLO4,fsul) were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The mutant lacked a low molecular weight (13,000) polypeptide present in the extract from the parent strain. This polypeptide difference was also observed when five other independently isolated non-flocculent mutants of the parent strain were analyzed. One of these five mutants was characterized genetically and also in this case was non-flocculence shown to be due to an unlinked suppressor mutation of FL04. This suppressor gene is designatedfsu2. Analysis of petites with different flocculation phenotypes further extended the correlation between non-flocculence and the absence of the polypeptide.The polypeptide was isolated by gel filtration in the presence of sodium dodecyl sulfate followed by ionexchange chromatography on DEAE-cellulose. By electrophoretic analysis the purity of the preparation was estimated to be 95%. From amino acid analysis the polypeptide was calculated to consist of 121 residues with a molecular weight of 12,900 daltons.Differential extraction of proteins iodinated in situ by lactoperoxidase suggested an external location of the polypeptide.