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Ascorbate and its interaction with plasma membrane redox systems

Martijn M. van Duijn

Ascorbate and its inter action with plasma membr ane redo x systems - Martijn M. va n Duijn

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Ascorbate and its interaction with plasma membrane redox systems

PROEFSCHRIFT

ter verkrijging van

de graad van Doctor aan de Universiteit Leiden, op gezag van de Rector Magnificus Dr. D.D. Breimer,

hoogleraar in de faculteit der Wiskunde en Natuurwetenschappen en die der Geneeskunde,

volgens besluit van het College voor Promoties te verdedigen op donderdag 31 mei 2001

te klokke 16.15 uur

door

Martijn Maurice van Duijn geboren te Katwijk

in 1972

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Promotiecommissie:

Promotor: Prof Dr H.J. Tanke

Co-promotores: Dr P.J.A. van den Broek Dr J. van der Zee

Referent: Prof Dr H. Goldenberg (University of Vienna, Austria)

Overige leden: Prof Dr G.W. Canters

Printed by PrintPartners Ipskamp ISBN 90-9014715-2

Note to the cover:

The redox systems described in this thesis form a gate in the plasma membrane

of cells, that allows electrons from ascorbate (vitamin C) to cross to the exterior

face of the cell. The cover symbolizes such a passageway in the membrane bilayer.

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"The great tragedy of science -- the slaying of a beautiful hypothesis by an ugly fact."

Thomas Huxley

Voor Judith

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Abbreviations

AFR ascorbate free radical Asc L-ascorbic acid

DHA dehydroascorbic acid

DiSC

3

(5) 3,3’-dipropylthiadecarbocyanine iodide DTPA diethylenetriaminepentaacetic acid EDTA ethylenediaminetetraacetic acid ESR electron spin resonance

FIC ferricyanide FOC ferrocyanide GSH glutathione

HPLC high performance liquid chromatography Ni(en)

32+

tris(ethylenediamine)nickel(II) chloride PBS phosphate buffered saline

p CMBS para -chloromerucribenzenesulfonic acid RDA recommended dietary allowance

ROS reactive oxygen species

TEMPO 2,2,6,6-tetramethylpiperidine-N-oxyl

TEMPOL 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl

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