Purification of chicken testicular müllerian inhibiting substance by ion exchange and high-performance liquid chromatography.

A highly purified protein molecule was obtained from the secretory proteins of 8-week-old chicken testes using ion-exchange column chromatographic procedures, including DEAE Bio-Gel A, CM Bio-Gel A, wheat germ lectin columns, and high-performance liquid chromatographic (hplc) separation techniques. This protein molecule has a molecular weight of 74,000 Da (74K protein). The isolated 74K protein induces regression of chicken Müllerian ducts grown in vitro. The 74K protein does not cause regression of cultured embryonic intestine or Wolffian duct. When the total testicular secretory proteins are resolved in a two-dimensional gel electrophoresis, approximately 120 polypeptides are obtained. The purified 74K protein has a pI of 6.1. Analysis of amino acid composition indicates that the 74K protein is relatively acidic in nature with a ratio of acidic to basic amino acids of 1.93.