Complete deficiency of glycophorin A in red blood cells from mice with targeted inactivation of the band 3 (AE1) gene

• Published in: Blood Vol. 91; no. 6; pp. 2146 - 2151
• Main Authors: HASSOUN, Y. H, HANADA, T, LUTCHMAN, M, SAHR, K. E, PALEK, J, HANSPAL, M, CHISHTI, A. H
• Format: Journal Article
• Language: English
• Published: Washington, DC The Americain Society of Hematology 15.03.1998
• Subjects: Animals; Anion Exchange Protein 1, Erythrocyte - deficiency; Anion Exchange Protein 1, Erythrocyte - genetics; Anion Exchange Protein 1, Erythrocyte - physiology; Biological and medical sciences; Biological Transport; Blood coagulation. Blood cells; Blood Proteins - analysis; Erythrocyte Membrane - metabolism; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods, hemostasis, fibrinolysis; Glycophorins - deficiency; Glycophorins - genetics; Glycophorins - metabolism; Membrane Proteins - metabolism; Mice; Mice, Knockout; Molecular and cellular biology; Polymerase Chain Reaction; RNA, Messenger - analysis; Transmembrane protein; Sialoglycoproteins; Vertebrata; Band 3 protein; Mammalia; Mouse; Animal; Glycophorine; Rodentia; Red blood cell
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.v91.6.2146

Glycophorin A is the major transmembrane sialoglycoprotein of red blood cells. It has been shown to contribute to the expression of the MN and Wright blood group antigens, to act as a receptor for the malaria parasite Plasmodium falciparum and Sendai virus, and along with the anion transporter, band 3, may contribute to the mechanical properties of the red blood cell membrane. Several lines of evidence suggest a close interaction between glycophorin A and band 3 during their biosynthesis. Recently, we have generated mice where the band 3 expression was completely eliminated by selective inactivation of the AE1 anion exchanger gene, thus allowing us to study the effect of band 3 on the expression of red blood cell membrane proteins. In this report, we show that the band 3 -/- red blood cells contain protein 4.1, adducin, dematin, p55, and glycophorin C. In contrast, the band 3 -/- red blood cells are completely devoid of glycophorin A (GPA), as assessed by Western blot and immunocytochemistry techniques, whereas the polymerase chain reaction (PCR) confirmed the presence of GPA mRNA. Pulse-label and pulse-chase experiments show that GPA is not incorporated in the membrane and is rapidly degraded in the cytoplasm. Based on these findings and other published evidence, we propose that band 3 plays a chaperone-like role, which is necessary for the recruitment of GPA to the red blood cell plasma membrane.