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Studies in the role of ubiquinone in the microbicidal events of neutrophils

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Call No.	QP95.8 .C73 1983
Author	Crawford, Dana Russell
Title	Studies in the role of ubiquinone in the microbicidal events of neutrophils
Date	1983
Abstract	The oxygen-dependent microbicidal events of neutrophils have been studied with reference to the triggering of the respiratory burst and with particular emphasis on the role of quinone. Ubiquinone as well as a previously described cytochrome b were found in neutrophils. the ubiquinone in human neutrophils was identified as ubiquinone-50, that in rats as ubiquinone-45. In human neutrophils, the ubiquinone and cytochrome b were located in a specific granule-like intracellular organelle. Several lines of evidence support the involvement of quinone in the respiratory burst of neutrophils. First, a number of short side chain quinones were found to stimulate burst activity in intact and in disrupted neutrophils. This stimulation exhibited saturation kinetics with respect to quinone concentration as would be expected for an enzyme mediated process. Second, a number of compounds structurally related to ubiquinone inhibited respiratory burst activity. Juglone, quercetin and nordihydroguaiaretic acid exhibited near micromolar K(,i) values. Third, the oxidation-reduction state of endogenous ubiquinone changed to one more oxidized following triggering of the burst by addition of latex. Furthermore, quercetin inhibited this oxidation-reduction change. The oxidative events of neutrophils were studied in greater detail using phagolysosomes isolated from latex-activated cells. These phagolysosomes were found to contain ubiquinone and to generate O(,2)('-) as measured by durohydroquinone-NBT reductase activity and NADPH-cytochrome c reductase activity. NADPH-cytochrome c reductase activity in phagolysosomes was greatly increased by the inclusion of phorbol myristate acetate during latex treatment. This reductase activity was inhibited by quercetin and nordihydroguaiaretic acid. NADPH-cytochrome c reductase activity and quinone content were enriched in phagolysosomes to a greater extent than marker enzymes for plasma membrane, azurophilic and specific granules, suggesting a role for a ubiquinone-containing, "tertiary" granule in the respiratory burst. The above results provide substantial evidence that ubiquinone is involved in the microbicidal events of neutrophils, presumably as a component of a novel electron transport chain that generates O(,2)('-).
Description	xiii, 99 leaves : ill.
Format	application/pdf
Subject	Ubiquinones. Neutrophils.
Issuing Body	Dartmouth Dissertations are made available through the Dartmouth Digital Library.
Degree	Thesis (Ph.D.)--Dartmouth College.
Copyright	All rights reserved. No part of this book may be reproduced in any form by any electronic or mechanical means without permission in writing from the author.
DOI	http://dx.doi.org/10.1349/ddlp.1495

Title	Page 1
Full Text	STUDIES ON THE ROLE OF UBIQUINONE IN THE MICROBICIDAL EVENTS OF NEUTROPHILS A Thesis Submitted to the Faculty in partial fulfillment of the requirements for the degree of Doctor of Philosophy by Dana R. Crawford DARTMOUTH COLLEGE Hanover, New Hampshire February, 1983

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