Fluorescence analysis of aldolase dissociation from the N-terminal of the cytoplasmic domain of band 3 induced by lanthanide.

The cytoplasmic domain of band 3 (CDB3) offers binding sites for several glycolytic enzymes and regulates the glycolysis of erythrocyte. The interaction between recombinant (His)(6)-tagged CDB3 and aldolase, one of the key enzymes that participated in erythrocyte glycolysis, was investigated in the presence of lanthanide. The results indicate that trace lanthanide blocks the inhibition of CDB3-(His)(6) to aldolase and leads to enhancement of aldolase activity. In agreement with activity studies, fluorescence spectra reveal that 4 microM lanthanum ions induce the complete dissociation of aldolase from the N-terminal of CDB3-(His)(6). Interestingly, the synchronous scanning fluorescence spectra of proteins in the presence of various concentrations of lanthanum ions suggest that the conformational change of CDB3-(His)(6) is significantly attributed to the alteration of tryptophan cluster microenvironment, while the aldolase conformation change is mainly derived from tyrosine microenvironment changes. Based on the observation that lanthanide ions induce the dissociation of aldolase from CDB3-(His)(6), it is suggested that the existence of trace lanthanide may affect the glycolysis of erythrocyte.