Structural Studies on the Leukocyte NADPH-Oxidase Generating Superoxide Oxyanion
Date
2014-01-15
Authors
Contributor
Advisor
Department
Instructor
Depositor
Speaker
Researcher
Consultant
Interviewer
Narrator
Transcriber
Annotator
Journal Title
Journal ISSN
Volume Title
Publisher
University of Hawaii at Manoa
Volume
Number/Issue
Starting Page
Ending Page
Alternative Title
Abstract
The membrane bound neutrophil NADPH-oxidase which catalyzes the one electron transfer from NADPH to molecular oxygen generating superoxide is a complex enzyme system made up of components whose structure and distribution in the membrane vary according to the system's state of activation. Cytochrome b558 is an important constituent of the NADPH-oxidase in neutrophils and B lymphocytes. Four components of the NADPH-oxidase have been identified: two membrane bound components, gp9 1 phox and p22phox, both subunits of cytochrome b558 ; and two cytosolic components, p47phox and p67phox (Chancock et al, 1994). The aim ofthis study was to gain further knowledge of the tertiary structure and arrangement of the large membrane bound component of cytochrome b558 (gp9 1 phox) in the bovine neutrophil plasma membranes during the resting state using proteolytic digestion with trypsin and papain to possibly identify the extra membranous components of cytochrome b558; i.e. In this neutrophil membrane preparation, the NADPH and FAD binding sites are postulated to be external to the membrane bilayer and should be exposed to proteolyticaction. Papaindigestion was also performed on octylglucoside solubilized and partially purified neutrophil membrane preparations. We will report on the superoxide-generating activity, NADPH oxidation and Iodonitrotetrazolium violet (INT) dye reduction profiles of membrane preparations and the correlation of such activities with proteolyticdigestiontime. Iodonitrotetrazolium violet (INT) is a dye which can be directly reduced by reduced flavins (FADH2). In addition to activity profiles, we will report on an analysis of the components of the membrane released by proteolysis.
Description
Keywords
Citation
Extent
39 pages
Format
Geographic Location
Time Period
Related To
Related To (URI)
Table of Contents
Rights
All UHM Honors Projects are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission from the copyright owner.
Rights Holder
Local Contexts
Collections
Email libraryada-l@lists.hawaii.edu if you need this content in ADA-compliant format.