Related Experiment Videos
Purification and characterization of insulin-like growth factor II (IGF II) and an IGF II variant from human placenta
F De Ceuninck1, J Willeput, M Corvol
1Institut National de la Santé et de la Recherche Médicale, Paris, France.
In order to purify variant IGF II peptides from human placenta, we have developed a purification procedure combining heparin affinity chromatography and cation-exchange, reversed-phase and size-exclusion HPLC. Two peptides were purified, both having apparent M(r) values of ca. 7300 Da as evaluated by SDS-PAGE. N-Terminal sequencing revealed IGF II and an IGF II variant in which Ser29 was replaced by the tetrapeptide Arg-Leu-Pro-Gly. The final yield of variant IGF II was about eight-fold lower than that of IGF II. Both pure peptides were functionally active as they bound to type I and type II IGF receptors from ovine and human placental membranes, as determined by crosslinking experiments and displacement curve studies.
In order to purify variant IGF II peptides from human placenta, we have developed a purification procedure combining heparin affinity chromatography and cation-exchange, reversed-phase and size-exclusion HPLC. Two peptides were purified, both having apparent M(r) values of ca. 7300 Da as evaluated by SDS-PAGE. N-Terminal sequencing revealed IGF II and an IGF II variant in which Ser29 was replaced by the tetrapeptide Arg-Leu-Pro-Gly. The final yield of variant IGF II was about eight-fold lower than that of IGF II. Both pure peptides were functionally active as they bound to type I and type II IGF receptors from ovine and human placental membranes, as determined by crosslinking experiments and displacement curve studies.