- Title Information
- Title
- Analysis of the Human Microtubule Interactome Using Multidimensional Chromatography and Tandem Mass Spectrometry
- Name:
Personal
- Name Part
- GONG, CHAO
- Role
- Role Term:
Text
- creator
- Origin Information
- Copyright Date
- 2013
- Physical Description
- Extent
- xix, 133 p.
- digitalOrigin
- born digital
- Note
- Thesis (Ph.D. -- Brown University (2013)
- Name:
Personal
- Name Part
- Bazemore-Walker, Carthene
- Role
- Role Term:
Text
- Director
- Name:
Personal
- Name Part
- Sun, Shouheng
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- Salomon, Arthur
- Role
- Role Term:
Text
- Reader
- Name:
Corporate
- Name Part
- Brown University. Chemistry
- Role
- Role Term:
Text
- sponsor
- Genre (aat)
- theses
- Subject
- Topic
- microtubule associated protein
- Subject
- Topic
- MAP
- Subject
- Topic
- two dimensional reversed phase HPLC
- Subject
- Topic
- peptide separation
- Subject
- Topic
- peptide diethylation
- Subject (FAST)
(authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/1020141")
- Topic
- Microtubules
- Subject (FAST)
(authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/1057576")
- Topic
- Peptides--Separation
- Record Information
- Record Content Source (marcorg)
- RPB
- Record Creation Date
(encoding="iso8601")
- 20131217
- Language
- Language Term:
Code (ISO639-2B)
- eng
- Language Term:
Text
- English
- Abstract
- Recent developments in mass spectrometer technology has increased the sensitivity and speed to an unprecedented level, along with the continuing improvements of high performance liquid chromatography (HPLC); they are combined as the most powerful tool for analysis of proteins, which are usually at low concentration levels in complicated biological samples. Often the challenge of shotgun proteomic analysis is related to the high complexity of the sample and limited instrument duty-cycle, which requires high resolution separation of components within the sample prior to mass spectrometer analysis. Aside from qualitative information of protein identifications from proteomic study, quantitative information is of increasing demand for the biological understanding of cellular functions or potential biomarkers.<br/>
We seek to investigate two dimensional reversed phase HPLC (2D-RP-HPLC) to provide high efficiency separation of peptides for proteomic study of complex biological systems. The utility of this method is illustrated for the characterization of the human microtubule interacting proteome, which is essential for the understanding of anti-tumor drug effects. In order to quantify expression changes of microtubule interacting proteins after drug treatment of cancer cells, we developed reductive diethylation strategy to isotopically label peptide amine groups for the differentiation of control samples and drug-treated samples.<br/>
Our results from the study of 2D-RP-HPLC peptide separation demonstrate its superior resolving power and selective separation mechanism for the comprehensive characterization of the microtubule interactome. Moreover, the reductive diethylation strategy exhibits high efficiency for stable isotopic labeling of peptides and no retention shift behavior of labeled peptide pairs even after 2D-RP-HPLC separation. Combining these two techniques provides an inexpensive and high-throughput workflow for quantitative proteomics. Finally, we identified a few protein candidates from the microtubule interactome that were up-regulated or down-regulated with response to a cancer therapeutic agent using this method.
- Identifier:
DOI
- 10.7301/Z0TT4P9M
- Access Condition:
rights statement
(href="http://rightsstatements.org/vocab/InC/1.0/")
- In Copyright
- Access Condition:
restriction on access
- Collection is open for research.
- Type of Resource (primo)
- dissertations