Weitreichender Elektronentransfer in biologischen Systemen

Graber, Michael. Weitreichender Elektronentransfer in biologischen Systemen. 2008, PhD Thesis, University of Basel, Faculty of Science.


Official URL: http://edoc.unibas.ch/diss/DissB_8360

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Ribonucleotide Reductases (RNR) are enzymes, which catalyze the reduction of all ribonucleotides
to their deoxyribonucleotides and therefore play an important role in DNA repair
und replication mechanisms. The class I RNR contains a stable diferric-tyrosyl radical cofactor
which is essential for the reduction of the substrates. This radical can directly be observed
by UV/Vis spectroscopy at 410 nm. After a substrate has bound in the active site, the radical
will be transferred along a conserved hydrogen bridge across a distance of 35 Å
The RNR’s proton coupled electron transfer (PCET) is in the focus of this thesis. The goal
was to gain insight in this process by stopped flow measurements and by laser flash photolysis
of caged substrates.
We could show that upon mixing cytidine diphosphate (CDP) and RNR no change of the tyrosyl
radical signal can be observed in millisecond scale. To examine faster processes in the
micro- and nanosecond scale, we synthesized two different caged substrates. We demonstrated
that upon irradiation, photocleavage takes place and CDP is released, which can be
converted to deoxycytidine diphosphate by RNR. After laser flash photolysis we observed no
change of the tyrosyl signal at λ = 410 nm. This might be due the fact that the PCET is even
faster than ~ 100 ns. Such a fast process is only in accordance with a hopping mechanism.
The goal of a second project was to build up a system to measure the speed of an electron
transfer in a double strand DNA-oligomer. We synthezised a modification based on thymidin,
which will be reduced to its radical anion upon laser irradiation at λ = 308 nm. An incorporated
porphyrin system acts as the electron acceptor and will be reduced to its radical anion.
By time dependent observation of the porphyrins absorption, the kET of this electron hopping
process can be measured.
We could synthesize both modifications (T* and porphyrin) and they were incorporated into a
double strand DNA-oligomer. We could show that upon irradiation of the porphyrin the chromophor
is stable against photochemical processes and no phosphorescence takes place.
Upon laser flash photolysis we observed a fast increase of the absorption at 445 nm followed
by a slow decrease. Fitting of the measured curves showed that the ET is faster than
kET ≥ 106 s-1 if only one A:T pair is between donor and acceptor. Future experiments with a
larger distance between thymidine and porphyrin will yield in more precise results concerning
the speed of the ET.
Advisors:Giese, Bernd
Committee Members:Wennemers, Helma
Faculties and Departments:05 Faculty of Science > Departement Chemie > Former Organization Units Chemistry > Bioorganische Chemie (Giese)
UniBasel Contributors:Giese, Bernd and Wennemers, Helma
Item Type:Thesis
Thesis Subtype:Doctoral Thesis
Thesis no:8360
Thesis status:Complete
Bibsysno:Link to catalogue
Number of Pages:91
Identification Number:
Last Modified:22 Jan 2018 15:50
Deposited On:13 Feb 2009 16:34

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