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Mechanism of protein aggregation leading to amyloid formation
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TitleInfo
Title
Mechanism of protein aggregation leading to amyloid formation
Name
(type = personal)
NamePart
(type = family)
He
NamePart
(type = given)
Xianglan
NamePart
(type = date)
1981-
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Xianglan He
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(authority = RULIB)
author
Name
(type = personal)
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(type = family)
Talaga
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(type = given)
David
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David Talaga
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Advisory Committee
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chair
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(type = family)
Levy
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(type = given)
Ronald
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Ronald Levy
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Advisory Committee
Role
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(authority = RULIB)
internal member
Name
(type = personal)
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(type = family)
Castner
NamePart
(type = given)
Edward
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Edward Castner
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Advisory Committee
Role
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internal member
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Ludescher
NamePart
(type = given)
Richard
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Richard Ludescher
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Advisory Committee
Role
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outside member
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Rutgers University
Role
RoleTerm
(authority = RULIB)
degree grantor
Name
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NamePart
Graduate School - New Brunswick
Role
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school
TypeOfResource
Text
Genre
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theses
Genre
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ETD doctoral
OriginInfo
DateCreated
(qualifier = exact)
2012
DateOther
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(type = degree)
2012-01
Place
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(type = code)
xx
Language
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(type = code)
eng
Subject
(authority = RUETD)
Topic
Chemistry and Chemical Biology
Subject
(authority = ETD-LCSH)
Topic
Amyloid beta-protein
Subject
(authority = ETD-LCSH)
Topic
Aggregation (Chemistry)
Subject
(authority = ETD-LCSH)
Topic
Amyloid
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Title
Rutgers University Electronic Theses and Dissertations
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ETD
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(type = host)
TitleInfo
Title
Graduate School - New Brunswick Electronic Theses and Dissertations
Identifier
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rucore19991600001
Identifier
ETD_3805
Identifier
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http://hdl.rutgers.edu/1782.1/rucore10001600001.ETD.000064219
Identifier
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doi:10.7282/T3MS3RS5
PhysicalDescription
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electronic resource
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application/pdf
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text/xml
Extent
xxiv, 152 p. : ill.
Note
(type = degree)
Ph.D.
Note
(type = bibliography)
Includes bibliographical references
Note
(type = vita)
Includes vita
Note
(type = statement of responsibility)
by Xianglan He
Location
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NjNbRU
Abstract
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Amyloid deposition has been observed in more than 20 diseases. Each amyloid-related disease has a particular precursor protein or peptide that converts from itsnative state to insoluble cross-βamyloid assemblies. Understanding the amyloid aggre-gation mechanism is the basis for development of a rational strategy for preventing theaggregation cascade events. This dissertation contributes to the elucidation of amyloidaggregation mechanism by characterizing the aggregation intermediates with comple-mentary biophysical techniques and clarifying the fluorescence mechanism of a standardamyloid probe, thioflavin T (ThT).
In the first part of this dissertation, aggregation state distribution was determinedat single-molecule level with atomic force microscopy (AFM), by development of aparticle size analysis package for AFM images. Combining the aggregation state dis-tribution from AFM and dynamic light scattering (DLS) measurements, a quantita-tive reservoir-nucleation model forβ-lactoglobulina(β-LGa) amyloid formation wasproposed based on kinetic simulation. The model successfully predicted the resultsof 1-anilino-8-naphthalene sulfonate (ANS) fluorescence and seeding experiments. Anaggregation free-energy landscape was constructed based on the simulation. A clas-sification scheme for oligomeric species was proposed to evaluate their roles during aggregation, based on their locations on the aggregation free-energy landscape. Dif-ferent types of oligomers were related to the amyloid cascade hypothesis and the toxicoligomer hypothesis for amyloid-related diseases.
The second part of this dissertation focuses on fluorescence mechanism of ThT,which is a standard tool for amyloid fibril detection and aggregation kinetic studies.DLS was used to probe the aggregation state of ThT in solutions of different condi-tions. The micelle model was disproved by showing the absence of ThT micelles insolution. The exciton model was disproved by fit of the concentration dependence ofThT fluorescence intensity to a fluorescent monomer model. Enhanced quantum yieldwas observed for surface-bound ThT in both bulk and single-molecule fluorescence mea-surements, with substantial shift in the excitation spectra, compared with fibril-boundThT. The presence of surface-bound ThT fluorescence calls for re-evaluation of reportedThT photophysical properties in solutions of low viscosity, which are likely dominatedby the surface effect. Similar lifetime features were observed for surface-bound andfibril-bound ThT. These results revealed that ThT bound to amyloid fibrils rigidly asit bound to the surface. Interaction of ThT withα-Synuclein (αSyn) fibrils was studiedwith single-molecule fluorescence microscopy. The single-molecule fluorescence polar-ization images showed that ThT bound toαSyn fibrils with its long axis parallel to thefibril axis.
In the first part of this dissertation, aggregation state distribution was determinedat single-molecule level with atomic force microscopy (AFM), by development of aparticle size analysis package for AFM images. Combining the aggregation state dis-tribution from AFM and dynamic light scattering (DLS) measurements, a quantita-tive reservoir-nucleation model forβ-lactoglobulina(β-LGa) amyloid formation wasproposed based on kinetic simulation. The model successfully predicted the resultsof 1-anilino-8-naphthalene sulfonate (ANS) fluorescence and seeding experiments. Anaggregation free-energy landscape was constructed based on the simulation. A clas-sification scheme for oligomeric species was proposed to evaluate their roles during aggregation, based on their locations on the aggregation free-energy landscape. Dif-ferent types of oligomers were related to the amyloid cascade hypothesis and the toxicoligomer hypothesis for amyloid-related diseases.
The second part of this dissertation focuses on fluorescence mechanism of ThT,which is a standard tool for amyloid fibril detection and aggregation kinetic studies.DLS was used to probe the aggregation state of ThT in solutions of different condi-tions. The micelle model was disproved by showing the absence of ThT micelles insolution. The exciton model was disproved by fit of the concentration dependence ofThT fluorescence intensity to a fluorescent monomer model. Enhanced quantum yieldwas observed for surface-bound ThT in both bulk and single-molecule fluorescence mea-surements, with substantial shift in the excitation spectra, compared with fibril-boundThT. The presence of surface-bound ThT fluorescence calls for re-evaluation of reportedThT photophysical properties in solutions of low viscosity, which are likely dominatedby the surface effect. Similar lifetime features were observed for surface-bound andfibril-bound ThT. These results revealed that ThT bound to amyloid fibrils rigidly asit bound to the surface. Interaction of ThT withα-Synuclein (αSyn) fibrils was studiedwith single-molecule fluorescence microscopy. The single-molecule fluorescence polar-ization images showed that ThT bound toαSyn fibrils with its long axis parallel to thefibril axis.
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Rights
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The author owns the copyright to this work.
RightsHolder
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Name
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He
GivenName
Xianglan
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Permission or license
DateTime
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2012-01-06 17:27:24
AssociatedEntity
Name
Xianglan He
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.
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2012-01-31
DateTime
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2013-01-30
Type
Embargo
Detail
Access to this PDF has been restricted at the author's request. It will be publicly available after January 30th, 2013.
Copyright
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Copyright protected
Availability
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Open
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Permission or license
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