PRELIMINARY INVESTIGATION ON THE FREE THIOLS AND DISULFIDE BONDS IN HUMAN HEPATOPOIETIN

摘要

rhHPO, the potential candidate for curing human liver diseases, has stirred the molecular biology and pathology. The recombinant product will be submitted to clinical trials very soon. Meanwhile, investigations on its advanced structure including disulfide bonds and conformations are going on. Some important results in the determination of free thiols and disulfide bonds of rhHPO using the strategy of partly reduced SDS-PAGE, vinylpyridine alkylation, Glu-C digestion and MALDI-TOF-MS are described in this paper. Two intrachain disulfide bonds and four free thiols exist in rhHPO monomer which forms dimer/polymer through at least one interchain disulfide bond. Direct locating of free thiols and disulfide bonds is in progress.