UV Raman evidence of a tyrosine in apo-human serum transferrin with a low pKa that is elevated upon binding of sulphate

Abstract

The binding of sulphate to human serum apo-transferrin has been examined by ultraviolet absorption and ultraviolet resonance Raman difference spectroscopies between pH 6.0 and 9.0. The ultraviolet absorption data reveals a negative feature at 245 nm that increases in magnitude with pH, with an apparent pKa of 7.57, which the Raman difference data reveals to be due to tyrosine. The pKa of this tyrosine is unusually low and is measured at 7.84 by the Raman difference method and is elevated to greater than 9.0 upon addition of sulphate. Previous studies on the N-lobe imply that Tyr 188 is the tyrosine with a low pKa and also that Arg 124 is the primary binding site for the sulphate. The functional relevance may be that with sulphate bound, both carbonate binding and the deprotonation of Tyr will be disfavoured, and as a result so is iron binding.