DARWIN Digitale Dissertationen German Version Strich

FU Berlin
Digitale Dissertation

Lorin Wehr :
Investigations to protein aggregation processes by means of dynamic and electrophoretic light scattering
Untersuchungen zu Proteinaggregationsprozessen mittels dynamischer und elektrophoretischer Lichtstreuung

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|Abstract| |Table of Contents| |More Information|

Abstract

The elucidation of the 3-dimensional structures of proteins needs crystals suitable for X-ray analysis. An absolute prerequisite for any crystallographic analysis is that well-ordered single crystals of the proteins are obtained. Unfortunately, the crystallisation of proteins and their complexes is still more of an art than a science and represents the major bottleneck in the determination of the 3d-structures by X-ray analysis. The present investigations provide an insight into molecular interactions, optimal crystallisation conditions and enhanced and novel crystallisation kits. For this photon-correlation-spectroscopy and laser-doppler-velocimetry were utilized. The former method provides essential information on the diffusion coefficient and hence on the radius and particle size distribution in the protein solutions. The latter method determines the zeta potential and the surface net charge of a protein via the measured electrophoretic mobility. Both measuring systems, also used in hanging drops with volumes of 10μl, provided a better understanding in aggregation phenomena, which could be partly correlated with optimal crystallisation conditions. The funfamental coincidence of the measured data with the aggregation state of the proteins in form of crystals or amorphous precipitate is confirmed, but a sure prediction between the outcome and the conditions of a protein crystallisation is only tendentiously possible. Nevertheless it is feasible to reduce the expense for a successful crystallisation by achieving additional information by means of light scattering techniques. The usefulness lies in the ability of getting a rational protocol of protein crystallisation and in the facilitation of protein crystallisation diagnostics.

Table of Contents

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0 Titelblatt und Inhaltsverzeichnis

1 Einleitung und Zielsetzung

2 Theoretischer Teil

2.1 Zur Physik der Kristallisation

2.2 Grundlagen aus der Kolloidchemie

2.3 Zur Theorie der Lichtstreuung

3 Experimenteller Teil

3.1 Meßmethoden und Datenerfassung

3.2 Materialien

3.3 Apparative Aufbauten

4 Meßergebnisse

4.1 Monomer-Monomer-Wechselwirkungen

4.2 Aggregationsprozesse

4.3 Stabilitätsübergänge

5 Diskussion der Ergebnisse

6 Zusammenfassung

7 Abstract

8 Anhang

8.1 Zur Datenauswertung

9 Persönliches

10 Literaturverzeichnis

More Information:

Online available: http://www.diss.fu-berlin.de/2002/140/indexe.html
Language of PhDThesis: german
Keywords: aggregation of proteins, electrolyte solutions, hanging drops, dynamic and electrophoretic light scattering
DNB-Sachgruppe: 30 Chemie
Date of disputation: 30-May-2002
PhDThesis from: Fachbereich Biologie, Chemie, Pharmazie, Freie Universität Berlin
First Referee: Prof. Dr. Wolfram Saenger
Second Referee: Prof.Dr. Peter Luger
Contact (Author): lorin.wehr@dlr.de
Contact (Advisor): saenger@chemie.fu-berlin.de
Date created:02-Aug-2002
Date available:05-Aug-2002

 

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