Limited and Pulse Proteolysis of Human Hemoglobin

Abstract

Using the title methods, the denaturation of human hemoglobin and its subunits was studied, the ultimate goal being to assess the applicability of both methods in research on pathogenic protein mutations. Pulse proteolysis affords a denaturation curve of hemoglobin; the mehod is easy to perform showing good repeatability and low sample consumption. Using the limited proteolysis followed by MALDI-TOF MS, we studied proteolytic resistance of fragments of α- and β-globin chains. Despite their nearly identical structure, the differences in amino acid compositions of both polypeptides affect the conformational stability of their tryptic products. Both methods are able to provide valuable information on protein stability, their main advantage being low cost and simplicity.