Modulation of erythrocyte deformability by PKC activity.
The interactions between membrane, peripheral and cytoskeleton proteins are responsible for the maintenance of erythrocyte deformability (EEI) and some of these interactions are modulated by PKC activity. Protein band 3 of the erythrocyte membrane is phosphorylated by phosphotyrosine kinases (PTK) and dephosphorylated by phosphotyrosine phosphatase (PTP). It was previously described by us a signal transduction mechanism that describes a possible pathway connecting an erythrocyte external membrane protein, acetylcholinesterase (AChE), with protein band 3. So how does PKC activity modulate EEI when protein band 3 is phosphorylated or dephosphorylated in absence or presence of AChE effectors?To answer this we used phorbol 12-myristate 13-acetate (PMA) as an activator and chelerythrine chloride as inhibitor of PKC and also band 3 modulators of band 3 phosphorylation degree, in presence and absence of AChE effectors in order to measure in whole blood samples EEI. Our results showed that erythrocyte deformability was significantly (i) decreased by inhibition of PKC, in absence and presence of AChE inhibitor velnacrine (ii) increased with PMA in absence and presence of ACh and (iii) decreased in presence of calpeptin in absence and presence of either chelerythrine or PMA. These results establish dependence between cytoskeleton proteins, PKC activity, band 3 phosphorylation degrees and EEI. Better understanding of those proteins interactions on transduction mechanisms might trigger possible targets for drug action that would modulate EEI.
de Oliveira
Sofia
S
Instituto de Bioquimica, Unidade Biologia Microvascular e Inflamacao, Instituto de Medicina Moecular, Faculdade de Medicina, Universidade de Lisboa, Lisboa, Portugal. sloliveira@fm.ul.pt
Silva-Herdade
Ana S
AS
Saldanha
Carlota
C
D000110:Acetylcholinesterase / Q000378:metabolism; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004907:Erythrocyte Deformability; D004912:Erythrocytes / Q000201:enzymology*; D018056:Hemorheology / Q000379:methods; D006801:Humans; D008954:Models, Biological; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D033081:Shear Strength; D015398:Signal Transduction; D013314:Stress, Mechanical
D001457:Anion Exchange Protein 1, Erythrocyte; D011493:Protein Kinase C; D000110:Acetylcholinesterase