IMMUNOLOGICAL STUDIES ON ADRENAL GLANDS .4. CHEMICAL STUDIES ON BOVINE THERMOSTABLE ADRENAL-SPECIFIC ANTIGEN

  • 1 January 1964
    • journal article
    • research article
    • Vol. 7  (6) , 605-+
Abstract
The thermostable ethanol-insoluble fraction of bovine adrenal tissue has been studied by physical and chemical procedures. Analysis in the ultracentrifuge revealed theee sedimenting boundaries in such preparations; the corrected and extrapolated sedimentation coefficients were 5.7, 2.7 and 1.9 S. Free electrophoresis at pH 8.6 showed four to seven compounds; the major boundaries had (negative) mobilities of 12.3, 10.0, 5.6 and 1.2 X 10-5 cm.2 volt-1 sec.-1. Light absorption studies revealed a maximum in optical density at a wavelength of 260 m[mu], indicative of nucleic acid. The extinction coefficient at 280 m[mu] was found to be 35.5 at a concentration of 1 g. per 100 ml. The same parameter at 540 m[mu] (with biuret reagent) was found to be 2.5. To elucidate the chemical nature of the previously reported adrenal- specific antigen in these heterogeneous preparations, degradation studies were made, followed by complement fixation, haemagglutination and precipitation methods to measure the remaining activity. The antigen was not affected by periodate, but was readily destroyed by chymotrypsin. This indicated that it is part of a polypeptide structure. The thermostable preparations also contain a non-adrenal-specific antigen. This second antigen was not destroyed by chymotrypsin, but neither was it affected by periodate, suggesting that a protein may be implicated here also.