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Preparation of Growth Hormone from a Rat Mammosomatotropic Pituitary Tumor¹

Departments of Biochemistry, St. Jude Children's Research Hospital and University of Tennessee, Memphis, Tennessee 38101

Rat growth hormone was extracted from an acetone powder of transplantable mammosomatotropic pituitary tumors using buffered ammonium sulphate. Active material, precipitated by increasing the ammonium sulphate concentration to 1.5 M, was fractionated first by diethylaminoethyl cellulose chromatography and then by preparative polyacrylamide gel electrophoresis. The yield of material in the purest fraction was 44 µg/gm wet weight of starting material. Tibia assays showed that the crude extract gave a 60-micron stimulation in cartilage width at a 2.0-mg dose level, whereas the purest growth hormone fraction gave a 53-micron stimulation at a 0.15-mg dose level. Analytical polyacrylamide gel electrophoresis at pH 9.5 indicated that the tumor growth hormone migrated more rapidly than growth hormone isolated from pituitaries; a single, but rather broad band was observed. The overall purification obtained was 12-fold.

¹ Supported in part by USPHS Research Grant No. AM-07375 from the Institute of Arthritic and Metabolic Diseases (USPHS), the National Science Foundation Grant No. GB-5743, the Damon Runyon Memorial Fund for Cancer Research Grant No. DRG-743, and the American Lebanese Syrian Associated Charities (ALSAC).

² W. E. G. acknowledges support by USPHS Training Grant No. 5-T01-CA-05176 from the National Cancer Institute.

Received 6/20/68. Accepted 10/23/68.