Modulation of red cell glycolysis: interactions between vertebrate hemoglobins and cytoplasmic domains of band 3 red cell membrane proteins

2004 ◽  
Vol 287 (2) ◽  
pp. R454-R464 ◽  
Author(s):  
Roy E. Weber ◽  
Wolfgang Voelter ◽  
Angela Fago ◽  
Hartmut Echner ◽  
Estela Campanella ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Binding Site ◽  
Protein Band ◽  
Band 3 ◽  
Glycolytic Enzyme ◽  
Pentose Phosphate ◽  
Dependent Manner ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Vital Functions

Several vital functions/physical characteristics of erythrocytes (including glycolysis, the pentose phosphate pathway, ion fluxes, and cellular deformability) display dependence on the state of hemoglobin oxygenation. The molecular mechanism proposed involves an interaction between deoxyhemoglobin and the cytoplasmic domain of the anion-exchange protein, band 3 (cdB3). Given that band 3 also binds to membrane proteins 4.1 and 4.2, several kinases, hemichromes, and integral membrane proteins, and at least three glycolytic enzymes, it has been suggested that the cdB3-deoxyhemoglobin interaction might modulate the pathways mediated by these associated proteins in an O2-dependent manner. We have investigated this mechanism by synthesizing 10-mer peptides corresponding to the NH2-terminal fragments of various vertebrate cdB3s, determining their effects on the oxygenation reactions of hemoglobins from the same and different species and examining binding of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase to the erythrocytic membrane of mouse erythrocytes. The cdB3 interaction is strongly dependent on pH and the number of negative and positive charges of the peptide and at the effector binding site, respectively. It lowers the O2association equilibrium constant of the deoxygenated (Tense) state of the hemoglobin and is inhibited by magnesium ions, which neutralize cdB3's charge and by 2,3-diphosphoglycerate, which competes for the cdB3-binding site. The interaction is stronger in humans (whose erythrocytes derive energy predominantly from glycolysis and exhibit higher buffering capacity) than in birds and ectothermic vertebrates (whose erythrocytes metabolize aerobically and are poorly buffered) and is insignificant in fish, suggesting that its role in the regulation of red cell glycolysis increased with phylogenetic development in vertebrates.

scholarly journals Characterization of glycolytic enzyme interactions with murine erythrocyte membranes in wild-type and membrane protein knockout mice

Blood ◽  
2008 ◽  
Vol 112 (9) ◽  
pp. 3900-3906 ◽  
Author(s):  
M. Estela Campanella ◽  
Haiyan Chu ◽  
Nancy J. Wandersee ◽  
Luanne L. Peters ◽  
Narla Mohandas ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Membrane Protein ◽  
Knockout Mice ◽  
Protein Band ◽  
Band 3 ◽  
Glycolytic Enzyme ◽  
Erythrocyte Membranes ◽  
Multienzyme Complexes ◽  
Human Erythrocyte Membranes ◽  
Ankyrin Protein

Previous research has shown that glycolytic enzymes (GEs) exist as multienzyme complexes on the inner surface of human erythrocyte membranes. Because GE binding sites have been mapped to sequences on the membrane protein, band 3, that are not conserved in other mammalian homologs, the question arose whether GEs can organize into complexes on other mammalian erythrocyte membranes. To address this, murine erythrocytes were stained with antibodies to glyceraldehyde-3-phosphate dehydrogenase, aldolase, phosphofructokinase, lactate dehydrogenase, and pyruvate kinase and analyzed by confocal microscopy. GEs were found to localize to the membrane in oxygenated erythrocytes but redistributed to the cytoplasm upon deoxygenation, as seen in human erythrocytes. To identify membrane proteins involved in GE assembly, erythrocytes from mice lacking each of the major erythrocyte membrane proteins were examined for GE localization. GEs from band 3 knockout mice were not membrane associated but distributed throughout the cytoplasm, regardless of erythrocyte oxygenation state. In contrast, erythrocytes from mice lacking α-spectrin, ankyrin, protein 4.2, protein 4.1, β-adducin, or dematin headpiece exhibited GEs bound to the membrane. These data suggest that oxygenation-dependent assembly of GEs on the membrane could be a general phenomenon of mammalian erythrocytes and that stability of these interactions depends primarily on band 3.

scholarly journals Selective phenylglyoxalation of functionally essential arginyl residues in the erythrocyte anion transport protein.

1983 ◽  
Vol 81 (4) ◽  
pp. 453-484 ◽  
Author(s):  
P J Bjerrum ◽  
J O Wieth ◽  
C L Borders
Keyword(s):  
Intracellular Ph ◽  
Covalent Binding ◽  
Chloride Concentration ◽  
Protein Band ◽  
Band 3 ◽  
Anion Transport ◽  
Transport Protein ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Arginyl Residues

The red cell anion transport protein, band 3, can be selectively modified with phenylglyoxal, which modifies arginyl residues (arg) in proteins, usually with a phenylglyoxal: arg stoichiometry of 2:1. Indiscriminate modification of all arg in red cell membrane proteins occurred rapidly when both extra- and intracellular pH were above 10. Selective modification of extracellularly exposed arg was achieved when ghosts with a neutral or acid intracellular pH were treated with phenylglyoxal in an alkaline medium. The rate and specificity of modification depend on the extracellular chloride concentration. At 165 mM chloride maximum transport inactivation was accompanied by the binding of four phenylglyoxals per band 3 molecule. After removal of extracellular chloride, maximum transport inhibition was accompanied by the incorporation of two phenylglyoxals per band 3, which suggests that transport function is inactivated by the modification of a single arg. After cleavage of band 3 with extracellular chymotrypsin, [14C]phenylglyoxal was located almost exclusively in a 35,000-dalton peptide. In contrast, the primary covalent binding site of the isothiocyanostilbenedisulfonates is a lysyl residue in the second cleavage product, a 65,000-dalton fragment. This finding supports the view that the transport region of band 3 is composed of strands from both chymotryptic fragments. The binding of phenylglyoxal and the stilbene inhibitors interfered with each other. The rate of phenylglyoxal binding was reduced by a reversibly binding stilbenedisulfonate (DNDS), and covalent binding of [3H]DIDS to phenylglyoxal-modified membranes was strongly delayed. At DIDS concentrations below 10 10 micrometers, only 50% of the band 3 molecules were labeled with [3H]-DIDS during 90 min at 38 degrees C, thereby demonstrating an interaction between binding of the two inhibitors to the protomers of the oligomeric band 3 molecules.

scholarly journals Insulin-induced glycosylphosphatidylinositol (GPI) binding to red cell membrane proteins

2002 ◽  
Vol 67 (12) ◽  
pp. 819-824
Author(s):  
Nenad Tomasevic ◽  
Milan Nikolic ◽  
Vesna Niketic
Keyword(s):  
Membrane Proteins ◽  
Cell Membrane ◽  
Molecular Mass ◽  
Band 3 ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Physiological Disorders ◽  
Cell Membrane Proteins

In this work GPI binding to membrane proteins from erythrocytes of insulinoma patients for whom prolonged hyperinsulinism and hypoglycemia were characteristic, as well as from normal erythrocytes incubated with supraphysiological concentrations of insulin were analyzed. In the RBCs from insulinoma patients, covalent GPI binding to red cell membrane proteins in the spectrin/ankyrin region, band 4.1 and two proteins of molecular mass of 115 and 110 kD was demonstrated. In erythrocytes incubated with insulin label was associated with band 4.1 and two proteins of molecular mass of 115 and 110 kD. Extraction studies showed that the 100-kD proteins are unrelated to band 3 since they were found in Triton- prepared cytoskeleton. To our knowledge this is the first demonstration of such a modification of red cell skeletal proteins, and the first demonstration of post-translation GPI binding to red cell skeletal proteins in response to insulin. A mechanism proposed for GPI binding to red cell skeletal proteins as well as the relevance of these results for physiological disorders that are characterized by hyperinsulinism are briefly discussed.

PAROXYSMAL NOCTURNAL HEMOGLOBINURIA IN CHILDHOOD AND ADOLESCENCE

PEDIATRICS ◽  
1967 ◽  
Vol 39 (5) ◽  
pp. 675-688
Author(s):  
Denis R. Miller ◽  
Robert L. Baehner ◽  
Louis K. Diamond
Keyword(s):  
Fetal Hemoglobin ◽  
Normal Activity ◽  
Glycolytic Enzyme ◽  
Clinical Severity ◽  
Childhood And Adolescence ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Severity Of The Disease ◽  
Increased Activity ◽  
Aldolase A

Two cases of PNH in adolescence and childhood are reported. The first presented at age 7½ years with aplastic anemia and improved after splenectomy performed at age 14. The second, a 15-year-old girl, presented with a Coombs-positive hemolytic anemia and has had a course complicated by multiple peripheral thromboses. The clinical and laboratory manifestations, complications, and certain therapeutic aspects of PNH are discussed. Anticoagulant therapy appears indicated in the presence of multiple thrombotic episodes. Erythrocyte metabolic studies revealed normal glycolysis, ATP stability, and GSH content in the cells of a child with a normal reticulocyte count. Mild elevations of glycolysis, noted in the child with a reticulocytosis, was ascribed to a younger mean red cell population since further elevations found in the "top" reticulocyte-rich layer after centrifugation. Heparin, the anticoagulant used in these studies, had no adverse effect on glycolysis but did inhibit hemolysis and minimize ATP instability when compared to cells suspended in defibrinated serum. Erythrocytes fractionated by centrifugation revealed increased glycolytic enzyme activities of hexokinase, G3PD, PGK, TPI, PK, LDH, G6PD, and 6PGD in the reticulocyte-rich layer. Normal, rather than increased activity of aldolase, a membrane enzyme, may reflect damage to the red cell membrane. PFK, known to be decreased in the erythrocyte of neonates, showed normal activity, but it was lowest in the reticulocyte-rich layer. Fetal hemoglobin was elevated in this layer. AChE deficiency and increased suceptibility to hydrogen perioxide and acid hemolysis confirmed previous reports and were most marked in the young cell layer. The level of increased glycolytic rates and enzyme activity, AChE deficiency, acid hemolysis and peroxide hemolysis were related to the clinical severity of the disease.

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