Biochemical characterization of human erythrocytes fractionated by counter-current distribution in aqueous polymer two-phase systems.
The fractionation of normal human erythrocytes by counter-current distribution (CCD) in charge-sensitive dextran-polyethylene glycol two-phase systems was confirmed and extended to red blood cells from heterozygous beta-thalassaemic patients. The differences between the distribution profiles of normal (homogeneous) and abnormal (heterogeneous) red blood cells reflect their different surface-charge properties. As suggested by the decline of membrane sialic acid released after neuraminidase treatment and the specific activities of two age-dependent enzymes (membrane acetylcholinesterase and intracellular pyruvate kinase) in the distribution profiles (from the left- to the right-hand side fractions), the fractionation seems to be according to red blood cell age. A constancy of the 2,3-bisphosphoglycerate level was observed in ageing red blood cells.
Pinilla
M
M
Departmento de Bioquímica y Biología Molecular, Universidad de Alcalá, Alcala de Henares, Madrid, Spain.
de la Fuente
J
J
García-Pérez
A I
AI
Jimeno
P
P
Sancho
P
P
Luque
J
J
D019794:2,3-Diphosphoglycerate; D000110:Acetylcholinesterase / Q000097:blood; D000328:Adult; D001731:Bisphosphoglycerate Mutase / Q000097:blood; D002469:Cell Separation / Q000379:methods*; D003377:Countercurrent Distribution; D004163:Diphosphoglyceric Acids / Q000097:blood; D004563:Electrochemistry; D004905:Erythrocyte Aging; D004909:Erythrocyte Indices; D004910:Erythrocyte Membrane / Q000737:chemistry; D004912:Erythrocytes / Q000737:chemistry / Q000166:cytology*; D006801:Humans; D019158:N-Acetylneuraminic Acid; D009439:Neuraminidase / Q000494:pharmacology; D011770:Pyruvate Kinase / Q000097:blood; D011773:Pyruvates / Q000097:blood; D019289:Pyruvic Acid; D012794:Sialic Acids / Q000097:blood; D014867:Water; D017086:beta-Thalassemia / Q000097:blood
D004163:Diphosphoglyceric Acids; D011773:Pyruvates; D012794:Sialic Acids; D014867:Water; D019794:2,3-Diphosphoglycerate; D019289:Pyruvic Acid; D011770:Pyruvate Kinase; D000110:Acetylcholinesterase; D009439:Neuraminidase; D001731:Bisphosphoglycerate Mutase; D019158:N-Acetylneuraminic Acid