Reversible binding of hemoglobin to band 3 constitutes the molecular switch that mediates O2 regulation of erythrocyte properties

Haiyan Chu, Mary M. McKenna, Nathan A. Krump, Suilan Zheng, Laurel Mendelsohn, Swee Lay Thein, Lisa J. Garrett, David M. Bodine and Philip S. Low

Key points

  • The reversible association of deoxyHb with band 3 acts as an O2-triggered molecular switch to regulate erythrocyte properties.

  • Transgenic mice lacking the deoxyHb site on band 3 fail to respond to changes in O2 with changes in erythrocyte properties.


Functional studies have shown that the oxygenation state of the erythrocyte regulates many important pathways, including glucose metabolism, membrane mechanical stability and cellular ATP release. Deoxyhemoglobin, but not oxyhemoglobin, binds avidly and reversibly to band 3, the major erythrocyte membrane protein. Because band 3 associates with multiple metabolic, solute transport, signal transduction and structural proteins, the hypothesis naturally arises that the O2-dependent regulation of erythrocyte properties might be mediated by the reversible association of deoxyHb with band 3. To explore whether the band 3-deoxyHb interaction constitutes a "molecular switch" for regulating erythrocyte biology, we have generated transgenic mice with mutations in the deoxyHb binding domain of band 3. One strain of mouse contains a "humanized" band 3 in which the N-terminal 45 residues of mouse band 3 are replaced by the homologous sequence from human band 3, including the normal human band 3 deoxyHb binding site. The second mouse contains the same substitution as the first, except the deoxyHb site on band 3 (residues 12-23) has been deleted. Comparison of these animals with wild type mice demonstrates that the following erythrocyte properties are controlled by the O2-dependent association of hemoglobin with band 3: i) assembly of a glycolytic enzyme complex on the erythrocyte membrane which is associated with a shift in glucose metabolism between the pentose phosphate pathway and glycolysis, ii) interaction of ankyrin with band 3 and the concomitant regulation of erythrocyte membrane stability, and iii) release of ATP from the red cell which has been linked to vasodilation.

  • Submitted January 12, 2016.
  • Accepted September 20, 2016.