title: Human erythrocyte membranes contain a cytochrome b561 that may be involved in extracellular ascorbate recycling.
abstract: Human erythrocytes contain an unidentified plasma membrane redox system that can reduce extracellular monodehydroascorbate by using intracellular ascorbate (Asc) as an electron donor. Here we show that human erythrocyte membranes contain a cytochrome b(561) (Cyt b(561)) and hypothesize that it may be responsible for this activity. Of three evolutionarily closely related Cyts b(561), immunoblots of human erythrocyte membranes showed only the duodenal cytochrome b(561) (DCytb) isoform. DCytb was also found in guinea pig erythrocyte membranes but not in erythrocyte membranes from the mouse or rat. Mouse erythrocytes lost a majority of the DCytb in the late erythroblast stage during erythropoiesis. Absorption spectroscopy showed that human erythrocyte membranes contain an Asc-reducible b-type Cyt having the same spectral characteristics as recombinant DCytb and biphasic reduction kinetics, similar to those of the chromaffin granule Cyt b(561). In contrast, mouse erythrocytes did not exhibit Asc-reducible b-type Cyt activity. Furthermore, in contrast to mouse erythrocytes, human erythrocytes much more effectively preserved extracellular Asc and transferred electrons from intracellular Asc to extracellular ferricyanide. These results suggest that the DCytb present in human erythrocytes may contribute to their ability to reduce extracellular monodehydroascorbate.
authors.affiliation: Department of Biochemistry, University of Nebraska, Lincoln, Nebraska 68588, USA.
authors.forename: Dan
authors.initials: D
authors.lastname: Su
authors.forename: James M
authors.initials: JM
authors.lastname: May
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Koury
authors.forename: Han
authors.initials: H
authors.lastname: Asard
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000378:metabolism*; D002460:Cell Line; D002837:Chromaffin Granules / Q000201:enzymology; D003573:Cytochrome b Group / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D003683:Dehydroascorbic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D004386:Duodenum / Q000201:enzymology; D004910:Erythrocyte Membrane / Q000201:enzymology*; D005110:Extracellular Space / Q000201:enzymology* / Q000378:metabolism; D006168:Guinea Pigs; D006801:Humans; D008247:Lysosomes / Q000201:enzymology; D008565:Membrane Proteins / Q000502:physiology; D051379:Mice; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000502:physiology; D051381:Rats
chemical_list: D003573:Cytochrome b Group; D008565:Membrane Proteins; C023646:cytochrome b561; C000820:semidehydroascorbic acid; D010088:Oxidoreductases; C020666:glutathione dehydrogenase (ascorbate); D001205:Ascorbic Acid; D003683:Dehydroascorbic Acid