Nobuhiro Takahashi, Toshiaki Isobe, Dominic M Desiderio, Nico M Nibbering

Proteomic Biology Using LC/MS

Large Scale Analysis of Cellular Dynamics and Function
By Nobuhiro Takahashi, Isobe Takahashi, Dominic M. Desiderio et al.

• Gebundenes Buch

Produktbeschreibung

One of the primary goals of current proteomic research is the description of the composition, dynamics, and connections of the multi-protein "modules" and "machineries" that perform a wide range of biological functions in the cell. This is one of the first books to focus on the dynamic aspect of proteomes. The book introduces proteomics to the newcomer, reviews the theoretical aspects of proteomics and its state-of-the art technologies, along with a number of biological applications using "classical" proteomic technology. The first chapter discusses how LC/MS technologies work with bioinformatics to search for the genome-database for large-scale and high-throughput protein analysis. The second chapter describes the applicability of those methodologies using the most recent applications to analyze the dynamicsof the cellular protein complexes and functions that are related to their formations and dissolutions. The book also presents a new concept, the "Dynamome," or the expressionof a comprehensive molecular set that participates in the whole dynamic process of a series of cellular events.

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Produktdetails

• Wiley-Interscience Series on Mass Spectrometry
• Verlag: Wiley & Sons
• Artikelnr. des Verlages: 14666258000
• 1. Auflage
• Seitenzahl: 264
• Erscheinungstermin: 2. November 2007
• Englisch
• Abmessung: 236mm x 165mm x 18mm
• Gewicht: 515g
• ISBN-13: 9780471662587
• ISBN-10: 0471662585
• Artikelnr.: 22744744

Autorenporträt

Nobuhiro Takahashi, PhD, is a Professor of Applied Biological Science in the Department of Agriculture at Tokyo University of Agriculture and Technology. Toshiaki Isobe, PhD, is a Professor of Biochemistry in the Department of Chemistry at Tokyo Metropolitan University.

Inhaltsangabe

Introduction. Chapter 1. Overview of Proteomics. What is the proteomics? 1
2 Proteomic analysis using two
dimensional gel electrophoresis (2DE) and mass spectrometry . 1
2
1 Protein separations by 2DE. 1
2
2 Development of the technologies for protein identification . 1
2
3 The protein identification based on gel separation and mass spectrometry. 1
3 Strategies for characterizing an entire proteome and understanding the proteome function. 1
3
1 Modification
specific proteomics. 1
3
2 Activity
based profiling. 1
3
3 Sub
cellular (Organelle) proteomics. 1
3
4 Machinery [complex (interaction)] proteomics. 1
3
5 Dynamic proteomics. Chapter 2. Proteomic Tools for Analysis of Cellular Dynamics. 2
1 LC
BASED PROTEOMICS TECHNOLOGIES.   2
1
1 LC system for peptide separation .   LC
MS interface.   ESI apparatus .   Micro
and nano
capillary columns.   Packing materials. 1D
LC system.   Application of 1D
LC
MS/MS to shotgun analysis of moderately complex protein mixtures. Experimental example 2
1. 2D LC
MS system. Experimental example 2
2.   2
1
2 Application of LC
MS methods to functional proteomics. Sub
cellular (organelle) using a cell
surface modification reagent and cell fractionation. Experimental example 2
3 . Modification proteomics.   Phosphorylation site mapping. Experimental example 2
4.   Glycosylation site mapping. Experimental example 2
5.   Ubiquitinated protein identification by using ubiquitin
specific antibody . Experimental example 2
6. 2
2 Development of quantitative proteomics. 2
2
1 Isotope labeling for quantitative analysis using MS. in vivo labeling. in vitro labeling. 2
2
2 Quantitation strategies for LC
MS analysis of isotope labeled peptide mixture and software for computer analysis. 2
2
3 Label free quantitation software. 2
2
4 Absolute quantitation. Method using stable isotope labeled reference peptides. Method without using internal standards. Chapter 3. Dynamics of Functional Cellular Machinery: From Statics to Dynamics in Proteomic Biology. From statics to dynamics in proteomic biology. 3
1 DYNAMIC ANALYSIS OF CELLULAR FUNCTION. 3
1
1 Strategy for dynamic analysis of cellular machineries (multi
protein complexes). Approach collecting time dependent data. Approach utilizing stage specific protein association. 3
1
2 Methods for the isolation of a cellular machinery/multi
protein complex. Affinity
tag purification. Affinity chromatography with an antibody
or a protein
immobilized column.   Immuno
affinity purification using antibody
fixed beads.   Experimental example 3
1.   Pull
down purification using immobilized protein beads.   Experimental example 3
2. 3
1
3 Cellular machinery (multi
protein complex). 3
2 Dynamics of Ribosome Biogenesis . 3
2
1 Snapshot Analyses of Preribosomal Particles in Yeast . 3
2
2 Snapshot Analyses of Preribosomal Particles in Mammals. Experimental example 3
3. 3
2
3 Quantitative (dynamic) analysis using isotope labeled reagents.   Experimental example 3
4. 3
2
4 Orthogonal Comparison of the Process of Ribosome Biogenesis. 3
3 Dynamic analyses of Sub
cellular structures. 3
3
1 Proteome dynamics of the nucleolus.

Rezensionen

"The book covers the topic of proteomics and mass spectrometry analysis well and does so at a reasonably advanced level. This volume would be suitable for advanced graduate students and for nonexpert scientists to learn about the application of mass spectrometry techniques for proteomic studies." ( Quarterly Review of Biology , December 2008)