Specificity and binding capacity of human blood serum for tetrahydropterins

Binding behaviour of labelled biopterin, 6,7-dimethylpterin, 2,4-diamino-6,7-dimethylpteridin, and of their 5,6,7,8-tetrahydro derivatives was studied with human serum proteins. Binding capacity of serum proteins is in the range of 5 x 10-10--10-3 M without saturation for all these pteridines. Preincubation or competition with tetrahydrobiopterin does not influence binding of tetrahydro-6,7-dimethylpterin in these concentrations. The bulk of pteridines is bound through unspecific adsorption. Only at concentrations of less than 0.8 x 10-8 M does high affinity binding seem to be possible, corresponding to about 300 pmol/g protein, whereas physiological biopterin concentration is near 2 x 10-8 M. The binding proteins are very sensitive to ageing and lose their capacity during purification, whereas unspecific binding to serum proteins is only weakly influenced by alteration of salt concentration, pH, or temperature. Attempts to partially purify the binding proteins by ion exchange, dextran gel (Sephadex G-200), or affinity chromatography, demonstrate a specificity of tetrahydropterins for the alpha2-macroglobulin fraction. Due to the high lability of this protein fraction and of pteridine binding, purification of a protein which specifically binds tetrahydrobiopterin was not achieved.