Isolation and partial sequence analysis of rat basic somatomedin

Abstract

Rat basic somatomedin (Sm) was prepared with an improved purification scheme from the pooled sera of Wistar Furth rats previously inoculated with cells from the pituitary tumor MStT/W15. Amicon hollow fiber diafiltration facilitated the processing of large batches of serum, eliminating the necessity of running smaller portions of material on gel filtration columns to achieve the same qualitative separation. The yield from this technique was excellent. Basic rat Sm was separated from a C3ades Arg component of complement by a narrow pH range isoelectric focusing step. Subsequent chromatography on carboxymethyl-cellulose and Sephadex resins was very efficient, providing a 100-fold purification, with a recovery of Sm activity of approximately 65%. The final product was judged to be pure by a variety of criteria. Structural analysis of this material has demonstrated that the amino-terminal sequences of rat basic Sm and human insulin-like growth factor I are strikingly similar, confirming the homology proposed earlier on the basis of receptor cross-reactivity.