A Mutant of Human Red Cell Pyruvate Kinase with High Affinity for Phosphoenolpyruvate

Enzyme ◽  
1974 ◽  
Vol 18 (1-2) ◽  
pp. 37-47 ◽  
Author(s):  
P. Boivin ◽  
C. Galand
Keyword(s):  
Pyruvate Kinase ◽  
Red Cell ◽  
High Affinity

scholarly journals Effect of oxalate and malonate on red cell metabolism

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1389-1393
Author(s):  
E Beutler ◽  
L Forman ◽  
C West
Keyword(s):  
Pyruvate Kinase ◽  
Cell Metabolism ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Red Cell ◽  
Acid Analogue ◽  
2,3 Dpg

The addition of oxalate to blood stored in Citrate-phosphate-dextrose (CPD) produces a marked improvement in 2,3-diphosphoglycerate (2,3-DPG) preservation; an increase in 2,3-DPG levels can also be documented in short-term incubation studies. Oxalate is a potent in vitro inhibitor of red cell lactate dehydrogenase, monophosphoglycerate mutase, and pyruvate kinase (PK). In the presence of fructose 1,6-diphosphate the latter inhibitory effect is competitive with phospho(enol)pyruvate (PEP). Determination of the levels of intermediate compounds in red cells incubated with oxalate suggest the presence of inhibition at the PK step, indicating that this is the site of oxalate action. Apparent inhibition at the glyceraldehyde phosphate dehydrogenase step is apparently due to an increase in the NADH/NAD ratio. Oxalate had no effect on the in vivo viability of rabbit red cells stored in CPD preservatives for 21 days. Greater understanding of the toxicity of oxalate is required before it can be considered suitable as a component of preservative media, but appreciation of the mechanism by which it affects 2,3-DPG levels may be important in design of other blood additives. Malonate, the 3-carbon dicarboxylic acid analogue of oxalate late did not inhibit pyruvate kinase nor affect 2,3-DPG levels.

Evidence that spectrin binds to macromolecular complexes on the inner surface of the red cell membrane

1980 ◽  
Vol 42 (1) ◽  
pp. 1-22 ◽  
Author(s):  
D. Litman ◽  
D.J. Hsu ◽  
V.T. Marchesi
Keyword(s):  
Cell Membrane ◽  
Blood Cell ◽  
Glycophorin A ◽  
Red Cell ◽  
Exposed Surface ◽  
Red Cell Membrane ◽  
Macromolecular Complexes ◽  
High Affinity ◽  
Cytoplasmic Surface ◽  
Inner Surface

Spectrin binds to a population of high-affinity sites on the exposed surface of inverted vesicles prepared from human red blood cell ghost membranes. Optimal spectrin binding requires the presence of monovalent salts but does not require calcium or magnesium. The band 2 subunit of spectrin, prepared in SDS, can also bind to vesicles, but isolated band 1 is inactive. Pre-incubation of inverted vesicles with antibodies directed against the cytoplasmic segment of band 3 or against bands 4.1-4.2 inhibits the binding of spectrin to the same vesicles. Antibodies against the cytoplasmic portion of glycophorin A have no effect. These results suggest that spectrin binds to a protein acceptor on the cytoplasmic surface of the red cell membrane which is close to the cytoplasmic segments of bands 3 and 4.1 and/or 4.2.

scholarly journals Coexistence of congenital red cell pyruvate kinase and band 3 deficiency

2004 ◽  
Vol 26 (4) ◽  
pp. 297-300 ◽  
Author(s):  
R. Branca ◽  
E. Costa ◽  
S. Rocha ◽  
H. Coelho ◽  
A. Quintanilha ◽  
...  
Keyword(s):  
Pyruvate Kinase ◽  
Band 3 ◽  
Red Cell

Red cell pyruvate kinase deficiency: 17 new mutations of the PK-LR gene

2005 ◽  
Vol 129 (6) ◽  
pp. 839-846 ◽  
Author(s):  
Elisa Fermo ◽  
Paola Bianchi ◽  
Laurent R. Chiarelli ◽  
Frederic Cotton ◽  
Cristina Vercellati ◽  
...  
Keyword(s):  
Pyruvate Kinase ◽  
Red Cell ◽  
Pyruvate Kinase Deficiency ◽  
New Mutations

scholarly journals Effect of oxalate and malonate on red cell metabolism

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1389-1393 ◽  
Author(s):  
E Beutler ◽  
L Forman ◽  
C West
Keyword(s):  
Pyruvate Kinase ◽  
Cell Metabolism ◽  
Inhibitory Effect ◽  
Red Cells ◽  
Red Cell ◽  
Acid Analogue ◽  
2,3 Dpg

Abstract The addition of oxalate to blood stored in Citrate-phosphate-dextrose (CPD) produces a marked improvement in 2,3-diphosphoglycerate (2,3-DPG) preservation; an increase in 2,3-DPG levels can also be documented in short-term incubation studies. Oxalate is a potent in vitro inhibitor of red cell lactate dehydrogenase, monophosphoglycerate mutase, and pyruvate kinase (PK). In the presence of fructose 1,6-diphosphate the latter inhibitory effect is competitive with phospho(enol)pyruvate (PEP). Determination of the levels of intermediate compounds in red cells incubated with oxalate suggest the presence of inhibition at the PK step, indicating that this is the site of oxalate action. Apparent inhibition at the glyceraldehyde phosphate dehydrogenase step is apparently due to an increase in the NADH/NAD ratio. Oxalate had no effect on the in vivo viability of rabbit red cells stored in CPD preservatives for 21 days. Greater understanding of the toxicity of oxalate is required before it can be considered suitable as a component of preservative media, but appreciation of the mechanism by which it affects 2,3-DPG levels may be important in design of other blood additives. Malonate, the 3-carbon dicarboxylic acid analogue of oxalate late did not inhibit pyruvate kinase nor affect 2,3-DPG levels.

scholarly journals Hemoglobin Crete (beta 129 ala leads to pro): a new high-affinity variant interacting with beta o -and delta beta o -thalassemia

Blood ◽  
1979 ◽  
Vol 54 (1) ◽  
pp. 54-63 ◽  
Author(s):  
A Maniatis ◽  
T Bousios ◽  
RL Nagel ◽  
T Balazs ◽  
Y Ueda ◽  
...  
Keyword(s):  
Cell Morphology ◽  
Clinical Picture ◽  
Oxygen Affinity ◽  
High Oxygen ◽  
Red Cell ◽  
High Affinity ◽  
Contact Sites ◽  
Erythroid Hyperplasia ◽  
Proline Substitution ◽  
High Oxygen Affinity

Abstract Hemoglobin Crete, beta129 (h7)ala leads to pro, is a new mutant hemoglobin (Hb) with high oxygen affinity that was discovered in a Greek family in various combinations with beta- and deltabeta- thalassemia. The propositus, who presented an unusual clinical picture of an “overcompensated” hemolytic state, with erythrocytosis, splenomegaly, abnormal red cell morphology, and marked erythroid hyperplasia, appeared doubly heterozygous for Hb Crete and deltabeta- thalassemia. His red cells contained 67% Hb Crete and 30% Hb F, and the combination of these two hemoglobins resulted in a blood P50O2 of 11.2 mm Hg. A brother with Hb Crete trait (38% Hb Crete, 56% Hb A, blood P50O2 23.0 mm Hg) did not have significant erythrocytosis. Purified Hb Crete was heat-unstable and exhibited a high oxygen affinity, and a normal Bohr effect. We postulate that the beta 129 proline substitution disrupts the H helix, perturbing nearby residues involved in alpha 1 beta 1 contact sites of the Hb tetramer.

Acquired Red Cell Pyruvate Kinase Deficiency in Leukemias and Related Disorders

Enzyme ◽  
1975 ◽  
Vol 19 (5-6) ◽  
pp. 294-299 ◽  
Author(s):  
P. Boivin ◽  
C. Galand ◽  
J. Hakim ◽  
A. Kahn
Keyword(s):  
Pyruvate Kinase ◽  
Red Cell ◽  
Pyruvate Kinase Deficiency

scholarly journals [3H]Ouabain binding and Na+, K+-ATPase in resealed human red cell ghosts.

1983 ◽  
Vol 81 (3) ◽  
pp. 401-420 ◽  
Author(s):  
D G Shoemaker ◽  
P K Lauf
Keyword(s):  
Adenylate Kinase ◽  
Kinase Activity ◽  
Kinase Inhibitor ◽  
Adenosine Diphosphate ◽  
Red Cell ◽  
Ouabain Binding ◽  
Intracellular Magnesium ◽  
High Affinity ◽  
Binding Rate ◽  
High Affinity Binding Site

The interaction of the cardiac glycoside [3H]ouabain with the Na+, K+ pump of resealed human erythrocyte ghosts was investigated. Binding of [3H]ouabain to high intracellular Na+ ghosts was studied in high extracellular Na+ media, a condition determined to produce maximal ouabain binding rates. Simultaneous examination of both the number of ouabain molecules bound per ghost and the corresponding inhibition of the Na+, K+-ATPase revealed that one molecule of [3H]ouabain inhibited one Na+, K+-ATPase complex. Intracellular magnesium or magnesium plus inorganic phosphate produced the lowest ouabain binding rate. Support of ouabain binding by adenosine diphosphate (ADP) was negligible, provided synthesis of adenosine triphosphate (ATP) through the residual adenylate kinase activity was prevented by the adenylate kinase inhibitor Ap5A. Uridine 5'-triphosphate (UTP) alone did not support ouabain binding after inhibition of the endogenous nucleoside diphosphokinase by trypan blue and depletion of residual ATP by the incorporation of hexokinase and glucose. ATP acting solely at the high-affinity binding site of the Na+, K+ pump (Km approximately 1 microM) promoted maximal [3H]ouabain binding rates. Failure of 5'-adenylyl-beta-gamma-imidophosphate (AMP-PNP) to stimulate significantly the rate of ouabain binding suggests that phosphorylation of the pump was required to expose the ouabain receptor.

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