Purification of somatomedin-C from human plasma: chemical and biological properties, partial sequence analysis, and relationship to other somatomedins.
Somatomedin-C (SM-C) was purified from Cohn fraction IV of human plasma by a series of steps which included cation-exchange chromatography, gel filtration, isoelectric focusing, and reverse-phase high-pressure liquid chromatography. The peptide isolated contained 10,142 units/mg, as judged by a radioimmunoassay, representing a 780,000-fold purification from native plasma. The isolated peptide was basic (pI 8.1-8.5) and was judged to be no less than 90% pure. The best fit for integral amino acids was obtained with 78 residues. Limited sequence data were obtained on the N terminus and on five fragments obtained by tryptic digestion after blocking the lysine residues. Three of these fragments and the N terminus could be aligned with portions of insulin-like growth factor I (IGF-I). Of the 25 residues so aligned, 22 were identical with IGF-I. In addition, two tryptic fragments were obtained which are not present in the sequence of IGF-I. SM-C and IGF-I produced identical curves of displacement in radioimmunoassay and radioreceptor assays for SM-C and in an insulin radioreceptor assay. The potencies of SM-C and IGF-I in these assays differed significantly from IGF-II, somatomedin-A, and several preparations of multiplication stimulating activity. At a concentration of 1 ng/mL, SM-C stimulated the progression of Balb/c 3T3 cells into DNA synthesis and when injected in vivo restored mitosis in lens epithelium of hypophysectomized frogs.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D006801:Humans; D007018:Hypopituitarism / Q000097:blood; D007334:Insulin-Like Growth Factor I; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D011863:Radioimmunoassay; D011869:Radioligand Assay; D013002:Somatomedins / Q000097:blood* / Q000302:isolation & purification; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D010446:Peptide Fragments; D013002:Somatomedins; D007334:Insulin-Like Growth Factor I