Bifunctional substrates of erythrocyte carbonic anhydrase. Enzyme-catalyzed hydration and hydrolysis of pyruvate esters

Red cell lysis in isotonic solutions containing NH4Cl, NH4HCO3, and a carbonic anhydrase enzyme inhibitor (acetazolamide) is a function of erythrocyte enzyme activity and permeability of cells to the inhibitor. Erythrocyte carbonic anhydrase activity is at least fivefold greater and acetazolamide permeability about tenfold less for adults than for newborns. In this setting, greater than 99.9% of red cells from adults can be hemolyzed at a time when greater than 25% of those from newborns remain intact. This easily applied method may be useful when antenatal diagnosis of hemoglobinopathies is otherwise precluded by contaimination with maternal erythrocytes. The feasibility of differential hemolysis via NH4Cl--HCO3-mediated, acetazolamide- modulated reactions is shown by the successful isolation of the few fetal-origin erythrocytes present in grossly nonbloody amniotic fluids and, in one instance, by approximately 3300-fold enrichment of apparently authentic fetal-origin red cells from the arm blood of a woman in her 18th wk of pregnancy.

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Catalytic versatility of erythrocyte carbonic anhydrase. IX. Kinetic studies of the enzyme-catalyzed hydrolysis of 3-pyridyl and nitro-3-pyridyl acetates

Biochemistry ◽

10.1021/bi00802a003 ◽

1971 ◽

Vol 10 (26) ◽

pp. 4843-4852 ◽

Cited By ~ 16

Author(s):

Y. Pocker ◽

N. Watamori

Keyword(s):

Carbonic Anhydrase ◽

Kinetic Studies ◽

Carbonic Anhydrase Ix ◽

Erythrocyte Carbonic Anhydrase ◽

Hydrolysis Of

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Enrichment of erythrocytes of fetal origin from adult-fetal blood mixtures via selective hemolysis of adult blood cells: an aid to antenatal diagnosis of hemoglobinopathies

Blood ◽

10.1182/blood.v47.6.883.883 ◽

1976 ◽

Vol 47 (6) ◽

pp. 883-897 ◽

Cited By ~ 40

Author(s):

SH Boyer ◽

AN Noyes ◽

ML Boyer

Keyword(s):

Carbonic Anhydrase ◽

Enzyme Inhibitor ◽

Antenatal Diagnosis ◽

Carbonic Anhydrase Activity ◽

Red Cells ◽

Fetal Origin ◽

Successful Isolation ◽

Erythrocyte Enzyme ◽

Erythrocyte Carbonic Anhydrase ◽

Carbonic Anhydrase Enzyme

Abstract Red cell lysis in isotonic solutions containing NH4Cl, NH4HCO3, and a carbonic anhydrase enzyme inhibitor (acetazolamide) is a function of erythrocyte enzyme activity and permeability of cells to the inhibitor. Erythrocyte carbonic anhydrase activity is at least fivefold greater and acetazolamide permeability about tenfold less for adults than for newborns. In this setting, greater than 99.9% of red cells from adults can be hemolyzed at a time when greater than 25% of those from newborns remain intact. This easily applied method may be useful when antenatal diagnosis of hemoglobinopathies is otherwise precluded by contaimination with maternal erythrocytes. The feasibility of differential hemolysis via NH4Cl--HCO3-mediated, acetazolamide- modulated reactions is shown by the successful isolation of the few fetal-origin erythrocytes present in grossly nonbloody amniotic fluids and, in one instance, by approximately 3300-fold enrichment of apparently authentic fetal-origin red cells from the arm blood of a woman in her 18th wk of pregnancy.

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