CHANGING A PROTEIN INTO A GENERALIZED ACYLATING REAGENT - REACTION OF NUCLEOPHILES WITH 3,5-DIBROMOSALICYL TRIMESYL-((LYS-BETA-82)-(LYS-BETA-82))-HEMOGLOBIN

The reaction of human hemoglobin A with trimesoyl tris(3,5-dibromosalicylate) produces the 3,5-dibromosalicyl ester of the bis(amide) of trimesic acid and the epsilon-amino groups of each of the Lys-82 residues of the two beta-subunits of hemoglobin (3,5-dibromosalicyl trimesyl-((Lys-beta-82)-(Lys-beta-82))-hemoglobin; DBST-Hb). This cross-linked protein containing a reactive ester site behaves as an acylating agent toward a variety of biochemically interesting nucleophiles. The products are acyl derivatives, as established by chromatographic and mass spectral analysis. The procedures demonstrate that the multifunctional reagent converts the protein into a reagent which is capable of directly generating bioconjugates of defined structure.