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A functional characterization of CGI-58
Descriptive
TitleInfo
Title
A functional characterization of CGI-58
Name
(type = personal)
NamePart
(type = family)
McMahon
NamePart
(type = given)
Derek B.
NamePart
(type = date)
1980-
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Derek McMahon
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(authority = RULIB)
author
Name
(type = personal)
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(type = family)
Brasaemle
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(type = given)
Dawn L
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Dawn L Brasaemle
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Advisory Committee
Role
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(authority = RULIB)
chair
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Storch
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(type = given)
Judith
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Judith Storch
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Advisory Committee
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internal member
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(type = personal)
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(type = family)
Watford
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(type = given)
Malcolm
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Malcolm Watford
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Advisory Committee
Role
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(authority = RULIB)
internal member
Name
(type = personal)
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(type = family)
Dixon
NamePart
(type = given)
Joseph
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Joseph Dixon
Affiliation
Advisory Committee
Role
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(authority = RULIB)
internal member
Name
(type = personal)
NamePart
(type = family)
Carman
NamePart
(type = given)
George
DisplayForm
George Carman
Affiliation
Advisory Committee
Role
RoleTerm
(authority = RULIB)
internal member
Name
(type = personal)
NamePart
(type = family)
Han
NamePart
(type = given)
Gil-Soo
DisplayForm
Gil-Soo Han
Affiliation
Advisory Committee
Role
RoleTerm
(authority = RULIB)
outside member
Name
(type = corporate)
NamePart
Rutgers University
Role
RoleTerm
(authority = RULIB)
degree grantor
Name
(type = corporate)
NamePart
Graduate School - New Brunswick
Role
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(authority = RULIB)
school
TypeOfResource
Text
Genre
(authority = marcgt)
theses
OriginInfo
DateCreated
(qualifier = exact)
2012
DateOther
(qualifier = exact);
(type = degree)
2012-10
CopyrightDate
(qualifier = exact)
2012
Place
PlaceTerm
(type = code)
xx
Language
LanguageTerm
(authority = ISO639-2b);
(type = code)
eng
Abstract
(type = abstract)
Mutations in the gene CGI-58/ABHD5 cause Chanarin-Dorfman Syndrome, a Neutral Lipid Storage Disorder (NLSD) where many cells and tissues, including human skin fibroblasts, store excessive triacylglycerol (TAG). The protein, CGI‐58, has been characterized in vitro as both a co‐activator of adipose triglyceride lipase (ATGL) and a lysophosphatidic acid acyltransferase (LPAAT). We hypothesized that CGI‐58 LPAAT activity is not necessary for co-activation of ATGL. This hypothesis was investigated through 3 specific aims: 1) to identify LPAAT active site residues, 2) to demonstrate that CGI‐58 lacking LPAAT activity can co-activate ATGL, and 3) to analyze the lipid composition of cultured NLSD fibroblasts relative to normal human skin fibroblasts. A molecular model of CGI‐58 was created to identify potential active site residues. In the model, the putative LPAAT active site residues H329 and D334 were not in close proximity, suggesting that they may not be active site residues. Recombinant H329A and D334A CGI‐58 variants, when purified from BL21(DE3) E. coli, showed higher levels of LPAAT activity than purified wild-type CGI‐58. LPAAT activity was linked to a protein contaminant, likely plsC, the endogenous E. coli LPAAT. The purification of recombinant CGI‐58 was optimized to reduce contaminant proteins. These new preparations lacked LPAAT activity, yet retained the ability to co‐activate ATGL. Additionally, extracts of Bl21(DE3) cells expressing GST-tagged CGI‐58 lacked LPAAT activity when plsC was removed by centrifugation. The previously observed LPAAT activity was due to a protein contaminant; thus, CGI‐58 lacks LPAAT activity and LPAAT activity is not necessary for the co-activation of ATGL. Additionally, using a protein‐lipid overlay, CGI‐58 bound to phosphatidylinositol 3-phosphate [PI(3)P] and phosphatidylinositol 5-phosphate [PI(5)P] but not lysophosphatidic acid [LPA], the LPAAT substrate. CGI‐58 binding of PI(3)P or PI(5)P does not alter co-activation of ATGL. Finally, CGI-58 variant H84R, found in humans with NLSDi, was expressed in cultured NLSD cells and studied in vitro. H84R CGI-58 failed to reduce accumulated TAG of NLSD fibroblasts, unlike unmodified CGI-58 or the H84A variant. Both H84 variants lacked the ability to co-activate ATGL in vitro. Thus, H84R CGI‐58 contributes to the NLSD phenotype by failing to co-activate ATGL.
Subject
(authority = RUETD)
Topic
Nutritional Sciences
RelatedItem
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TitleInfo
Title
Rutgers University Electronic Theses and Dissertations
Identifier
(type = RULIB)
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Identifier
ETD_4304
PhysicalDescription
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(authority = gmd)
electronic resource
InternetMediaType
application/pdf
InternetMediaType
text/xml
Extent
xii, 122 p. : ill.
Note
(type = degree)
Ph.D.
Note
(type = bibliography)
Includes bibliographical references
Note
(type = statement of responsibility)
by Derek B. McMahon
Subject
(authority = ETD-LCSH)
Topic
Mutation (Biology)
Subject
(authority = ETD-LCSH)
Topic
Lipidoses
Identifier
(type = hdl)
http://hdl.rutgers.edu/1782.1/rucore10001600001.ETD.000066910
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TitleInfo
Title
Graduate School - New Brunswick Electronic Theses and Dissertations
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rucore19991600001
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(displayLabel = Rutgers, The State University of New Jersey)
NjNbRU
Identifier
(type = doi)
doi:10.7282/T34F1PGR
Genre
(authority = ExL-Esploro)
ETD doctoral
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Rights
RightsDeclaration
(ID = rulibRdec0006)
The author owns the copyright to this work.
RightsHolder
(type = personal)
Name
FamilyName
McMahon
GivenName
Derek
Role
Copyright Holder
RightsEvent
Type
Permission or license
DateTime
(encoding = w3cdtf);
(qualifier = exact);
(point = start)
2012-09-27 11:28:12
AssociatedEntity
Name
Derek McMahon
Role
Copyright holder
Affiliation
Rutgers University. Graduate School - New Brunswick
AssociatedObject
Type
License
Name
Author Agreement License
Detail
I hereby grant to the Rutgers University Libraries and to my school the non-exclusive right to archive, reproduce and distribute my thesis or dissertation, in whole or in part, and/or my abstract, in whole or in part, in and from an electronic format, subject to the release date subsequently stipulated in this submittal form and approved by my school. I represent and stipulate that the thesis or dissertation and its abstract are my original work, that they do not infringe or violate any rights of others, and that I make these grants as the sole owner of the rights to my thesis or dissertation and its abstract. I represent that I have obtained written permissions, when necessary, from the owner(s) of each third party copyrighted matter to be included in my thesis or dissertation and will supply copies of such upon request by my school. I acknowledge that RU ETD and my school will not distribute my thesis or dissertation or its abstract if, in their reasonable judgment, they believe all such rights have not been secured. I acknowledge that I retain ownership rights to the copyright of my work. I also retain the right to use all or part of this thesis or dissertation in future works, such as articles or books.
Copyright
Status
Copyright protected
Availability
Status
Open
Reason
Permission or license
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