Characterisation of oxidized recombinant human galectin-1

Abstract

Oxidized human galectin-1 plays a role in the immune response to injured axons. Over-expression of galectin-1 by cancer, in combination with cancer associated oxidative stress suggests oxidized human galectin-1 may also play a role(s) in tumourigenesis. Here we generate milligram quantities of oxidized human galectin-1 and undertake biophysical characterization. The protein adopts a number of different states. Two separable oxidized forms are identified that exist as largely mono-disperse solutions at higher milligram/ml concentrations. The presence of disulphide bonds is confirmed for these two protein forms, as is their change in overall shape and loss of lectin activity. Our studies lead to production of a particular mono-disperse oxidized human galectin-1 species that is anticipated most optimal for investigations requiring milligram/ml concentrations such as X-ray crystallography.