1. Status report on the study of biological systems at ISOLDE: IS448 and IS488
Monika Stachura, Lars Hemmingsen
University of Copenhagen, Denmark

2. Members of the collaboration
University of Copenhagen:
Lars Hemmingsen, Peter W. Thulstrup, Marianne L. Jensen, Monika Stachura, Vaida Arcišauskaitė, Søren T. Heide
University of Michigan:
Vincent L. Pecoraro, Saumen Chakraborty
University of Zürich:
Roland Siegel, Eva Freisinger, Silke Johansen,
Miquel B. Oliver
University of Szeged:
Bela Gyurcsik, Attila Jancso, Dániel Szunyogh
University of Wroclaw:
Marek Luczkowski
University of Münster:
Jens Mueller
University of Greifswald:
Sabrina Kargoll
ISOLDE/CERN:
Alexander Gottberg, Karl Johnston
20 participants
9 nationalities
8 institutes
6 countries
Monika Stachura INTC meeting July 2011

3. Summary of runs 2009 – 2011 Isotopes: 204mPb, 199mHg, 111Ag
Shifts: (done) + 6 (2011)‏ + 7 (bio-beta-NMR)
Experiments: ~75
Publications: 5 (+ 3 manuscripts in preparation)
Year IS IS488
mPb – 4 shifts (cancelled) mHg – 6 shifts (50 experiments)‏
111Ag – 3 shifts (0 experiments)‏
mHg – 3 shifts (23 experiments)‏
mHg – 4 shifts (October) Ag – 2 shifts (July)‏
Monika Stachura INTC meeting July 2011

4. Experiments Implantation into ice (GLM)‏
2009: 40 – 60 min
2010: 10 – 15 min
Sample preparation (bld. 115, OFFLINE!)‏
~10 min preparation
~10 min incubation
Measurements (two PAC instruments)‏
~ 3 hours
Data analysis (CERN, Copenhagen)‏
Monika Stachura INTC meeting July 2011

5. Samples Finished Small model complexes De novo designed peptides
~20 different compounds
Isotope: 204mPb, 199mHg
Finished
De novo designed peptides
~10 different peptides
Isotopes: 204mPb, 199mHg
Experimental conditions: pH
temperature
metal ion to peptide ratio
addition of other ions
different buffers
Natural proteins
HAH1, metallothionein, Ncp7, Lys
plants
Isotope: 199mHg
Nucleic acids
DNA, RNA
Isotope: 199mHg
Monika Stachura INTC meeting July 2011

6. Results Small model complexes De novo designed peptides
Hg(PhS)2
Hg(salen)‏
Small model complexes
to test the quality of DFT calculations of EFG
structural interpretation of more complex systems
Manuscript in preparation
De novo designed peptides
synthesized by V. L. Pecoraro
to study different coordination geometries of Hg(II)‏
Iranzo et al., J. Am. Chem. Soc. 2011, 133, 239­251
Chakraborty et al., Angew. Chem. Int. Ed. 2011, 50, 2049­2053
Manuscript in preparation
Monika Stachura INTC meeting July 2011

7. Natural proteins – HAH1 (Copenhagen & Michigan & Wroclaw)‏
A. K. Wernimont et al., Nature Structural Biology, 7, (2000)
Manuscript in preparation
Monika Stachura INTC meeting July 2011

8. Bio-beta NMR (I88)‏ Na+, K+, Mg2+, Ca2+, Zn2+,
Fe, Co, Ni, Cu, Mn, Mo, Cr(a)‏
(a)charge not given since this varies with oxidation state
Monika Stachura INTC meeting July 2011

9. Future projects and experiments
PAC Spectroscopy:
Studies of heavy metal ion binding to proteins and nucleic acids (RNA/DNA)‏
Water purification by bacteria
Heavy metal ion binding by plants and bacteria (in vivo experiments)‏
Record the first clearcut 204mPb PAC for proteins
Bio-beta-NMR Spectroscopy:
Setup optimization
Experiments with proteins
Shutdown period:
Shorter shutdown preferable (no beam in 2013, restart in May 2014)‏
Monika Stachura INTC meeting July 2011

10. Thank you ! Acknowledgements
A. Gottberg, K. Johnston, M. Kowalska, Y. Blumenfeld, J. G. Correia
The ISOLDE collaboration
ENSAR
Danish Research Council for Nature and Universe
Thank you !
Monika Stachura INTC meeting July 2011