The initial noncovalent binding of glucose to human hemoglobin in nonenzymatic glycation.

Mechanisms for nonenzymatic protein glycation have been extensively studied albeit with an emphasis at the later stages that gives rise to advanced glycation end products. No detailed investigation of the initial, noncovalent binding of d-glucose to human hemoglobin A (HbA) exists in the literature. Although anionic molecules 2,3-bisphosphoglycerate (BPG), inorganic phosphate (Pi) and HCO3(-) have been implicated in the latter stages of glycation, their involvement at the initial binding of glucose to HbA has not yet been assessed. Results from this computational study involving crystal structures of HbA predict that the transient, ring-opened glucose isomer, assumed to be critical in the later stages of glycation, is not directly involved in initial binding to the β-chain of HbA. All the five structures of glucose generated upon mutorotation will undergo reversible, competitive and slow binding at multiple amino acid residues. The ring-opened structure is most likely generated from previously bound pyranoses that undergo mutarotation while bound. BPG, Pi and HCO3(-) also reversibly bind to HbA with similar energies as glucose isomers (~3-5 kcal/mol) and share common binding sites with glucose isomers. However, there was modest amino acid residue selectivity for binding of certain anionic molecules (1-3 regions) but limited selectivity for glucose structures (≥ 7 regions). The clinical difference between average blood glucose and predicted HbA1c, and the presence of unstable HbA-glucose complexes may be more fully explained by initial noncovalent binding interactions and different concentrations of BPG, Pi and HCO3(-) in serum vs. erythrocytes.