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Mechanism of Na,K-ATPase decline during sheep red cell maturation

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:5904602
;

Na,K-ATPase of immature and mature sheep red cells of both the high-K/sup +/ and low-K/sup +/ genotypes as well as cells of both types matured in vitro was detected using polyclonal antiserum to sheep kidney Na,K-ATPase. Following SDS-PAGE and immunoblotting, the major reactive component was the approx. 100 kDa catalytic ..cap alpha.. subunit. A less prominent band migrating as a sharper, lower molecular weight (50 kDa) component than the kidney Na,K-ATPase ..beta.. subunit is apparent in reticulocytes but not mature cells. Membranes from both genotypes showed identical immunologically reactive peptides, except for the lower intensity of the ..cap alpha.. subunit in the mature cells of the low- compared to high-K/sup +/ sheep. Following culture of both types, moderate reduction in reactivity was apparent. Immunologically reactive ..cap alpha.. subunit as well as the 50 kDa species were detected in membranous material shed into the culture medium. This material was functionally inactive (lack of both (/sup 3/H) ouabain binding and Na/sup +/-dependent phosphorylation of Na,K-ATPase). The existence in reticulocytes of an intracellular pool of ouabain binding sites is evidenced in appearance of extra sites following rapid ATP depletion and also after addition of chloroquine. Taken together, these findings are consistent with a maturation-associated decrease of sodium pumps by a process of membrane recycling, processing and, to some extent, exocytosis.

Research Organization:
Montreal General Hospital Research Institute, Quebec
OSTI ID:
5904602
Report Number(s):
CONF-870644-
Journal Information:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 46:6; ISSN FEPRA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACID ANHYDRASES
ALKALI METALS
ANIMALS
ATP-ASE
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY
BODY FLUIDS
CARBOHYDRATES
CARDIAC GLYCOSIDES
CARDIOTONICS
CARDIOVASCULAR AGENTS
CELL CONSTITUENTS
CELL DIFFERENTIATION
CELL MEMBRANES
DOMESTIC ANIMALS
DRUGS
ELECTROPHORESIS
ELEMENTS
ENZYME ACTIVITY
ENZYMES
ERYTHROCYTES
GLYCOSIDES
HYDROLASES
IMMUNE REACTIONS
ISOTOPE APPLICATIONS
KIDNEYS
KINETICS
LABELLED COMPOUNDS
MAMMALS
MATERIALS
MEMBRANE PROTEINS
MEMBRANES
METALS
MOLECULAR WEIGHT
ORGANIC COMPOUNDS
ORGANS
OUABAIN
PHOSPHOHYDROLASES
POTASSIUM
PROTEINS
REACTION KINETICS
RECEPTORS
RETICULOCYTES
RUMINANTS
SHEEP
SODIUM
STROPHANTHINS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES