Purification and characterization of progenipoietins produced in Escherichia. coli.

摘要: The progenipoietins (ProGPs) are a family of genetically engineered chimeric proteins that contain receptor agonist activity for both fetal liver tyrosine kinase-3 and the granulocyte colony-stimulating factor receptor. These unique have previously been shown to induce proliferation multiple cell lineages. characterization two progenipoietins, ProGP-1 ProGP-4, refolded purified from an Escherichia coli expression system is described. ProGP molecules differ in orientation agonists and, addition, ProGP-4 contains has circularly permuted modulate its activity. Static light scattering analyses demonstrated exist as dimers, most likely through non-covalent interaction domains. comparable secondary structures, analyzed by circular dichroism; however, their tertiary measured intrinsic fluorescence, were be different. Differential scanning calorimetry thermal stability these was indistinguishable. Interestingly, dual yielded only single melting temperature value intermediate between individual components, indicating behave domain protein during denaturation. This study describes purification physico-chemical properties this class generated using E. system.