Investigations of enzymatic reactions using mass spectrometry
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Contents
Contents
Investigations of enzymatic reactions using mass spectrometry
Gunter Kuhnle
Date:
8. Juni 2001
Contents
List of Figures
List of Tables
Introduction
Mass spectrometry
Development of mass spectrometry
Mass spectrometric techniques
Ion sources
Mass analysers
Ribonuclease T1: a model enzyme
Reaction mechanism
Specificity of RNase T1
Protein design and specificity alteration
Kinetic investigations of RNase T1
Enzymatic reactions
Kinetic investigations of enzymatic reactions
Conceptual formulation
Experimental
Materials
Chemicals and Solvents
Buffers
Substrates and standards
Enzymes
Matrix and Solvents
Instrumentation
Mass spectrometry
HPLC and FIA
nano
ES ion source
Other technical equipment
Software
Data acquisition
Data processing
Methods
Sample preparation
Mass spectrometry
Electrospray mass spectrometry
nano
ES mass spectrometry
MALDI-TOF-MS
Results and Discussion
Off-line reaction monitoring
Reaction assay
Data acquisition and processing
Data processing with
kassandra
Quantification
Determination of kinetic parameters
5',3'-GpC transesterification
5',3'-ApC transesterification
Hydrolysis of the cyclic 2',3'-phosphodiester
Conclusions
On-line reaction monitoring using
nano
ES
Development of a
nano
ES ion source
Construction and Design
Enzymatic hydrolysis of 5',3'-GpC by RNase T1
Multiple substrate monitoring
Conclusions
Cleavage pattern analysis
Substrate design
Analysis of cleavage pattern
RNase T1 wild type
RNase T1
RV
Conclusions
Summary
Bibliography
Abbreviations
General abbreviations
Nucleotides
Amino acids
Publications resulting from this work
Publications
Presentations at Conferences
Curriculum vitae
Acknowledgments
Selbständigkeitserklärung
About this document ...
Gunter Kuhnle 2001-06-04
