Ascorbate is regenerated by HL-60 cells through the transplasmalemma redox system

• Published in: Biochimica et biophysica acta Vol. 1073; no. 2; pp. 380 - 385
• Main Authors: Alcain, Francisco J., Buron, M. Isabel, Villalba, Jose M., Navas, Placido
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 04.03.1991; Elsevier
• Subjects: Ascorbate; Ascorbic Acid - metabolism; Ascorbic Acid - pharmacology; Biological and medical sciences; Cell Count; Cell Division - drug effects; Cell Line; Cell Membrane - metabolism; Cell membranes. Ionic channels. Membrane pores; Cell structures and functions; Concanavalin A - pharmacology; Dehydroascorbic Acid - metabolism; Ferricyanides - metabolism; Free Radicals; Fundamental and applied biological sciences. Psychology; HL-60; Humans; Kinetics; Lactates - metabolism; Lactic Acid; Molecular and cellular biology; NAD - metabolism; NADH, NADPH Oxidoreductases - metabolism; Oxidation-Reduction; Oxygen Consumption; Plasma membrane; Redox system; Wheat Germ Agglutinins - pharmacology; Ascorbate; HL-60; Redox system; AFR; Plasma membrane; Cell culture; Regeneration; Ascorbic acid derivatives; Enzymatic activity; Kinetics; Electron microscopy; Morphometry; Metabolism
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/0304-4165(91)90146-8

Ascorbate was maintained in the media during a long-term culture by HL-60 cells. The chemical oxidation of ascorbate was reversed in vitro by living HL-60 cells and was related to the amount of cells added. The increase of NADH concentration by lactate addition to cells was accompanied by an increase of both ascorbate regeneration and ferricyanide reduction. Further, plasma membrane enriched fractions from HL-60 cells revealed enhancement of both ascorbate regeneration and ferricyanide reduction in the presence of NADH when previously treated with detergent. The blockage of cell surface carbohydrates by wheat germ agglutinin (WGA) and Concanavalina ensiformis (Con A) lectins significantly inhibited the regeneration of ascorbate caused by the cells. These results support the idea that ascorbate is externally regenerated by the NADH-ascorbate free radical reductase as a part of the transplama membrane redox system.