Modulation of Erythrocyte Membrane Mechanical Function by β-Spectrin Phosphorylation and Dephosphorylation (∗)

• Published in: The Journal of biological chemistry 1995-03, Vol.270 (10), p.5659-5665
• Main Authors: Manno, Sumie, Takakuwa, Yuichi, Nagao, Kaoru, Mohandas, Narla
• Format: Article
• Language: English
• Subjects: Actins - blood; Adenosine Triphosphate - pharmacology; Erythrocyte Membrane - drug effects; Erythrocyte Membrane - physiology; Humans; Kinetics; Luminescent Measurements; Macromolecular Substances; Membrane Proteins - blood; Membrane Proteins - isolation & purification; Phosphoproteins - blood; Phosphoproteins - isolation & purification; Phosphorylation; Protein Kinases - blood; Spectrin - isolation & purification; Spectrin - metabolism; Vanadates - pharmacology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.270.10.5659

The mechanical properties of human erythrocyte membrane are largely regulated by submembranous protein skeleton whose principal components are α- and β-spectrin, actin, protein 4.1, adducin, and dematin. All of these proteins, except for actin, are phosphorylated by various kinases present in the erythrocyte. In vitro studies with purified skeletal proteins and various kinases has shown that while phosphorylation of these proteins can modify some of the binary and ternary protein interactions, it has no effect on certain other interactions between these proteins. Most importantly, at present there is no direct evidence that phosphorylation of skeletal protein(s) alters the function of the intact membrane. To explore this critical issue, we have developed experimental strategies to determine the functional consequences of phosphorylation of βspectrin on mechanical properties of intact erythrocyte membrane. We have been able to document that membrane mechanical stability is exquisitely regulated by phosphorylation of β-spectrin by membrane-bound casein kinase I. Increased phosphorylation of β-spectrin decreases membrane mechanical stability while decreased phosphorylation increases membrane mechanical stability. Our data for the first time demonstrate that phosphorylation of a skeletal protein in situ can modulate physiological function of native erythrocyte membrane.