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

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

Summary of runs 2009 – 2011 Isotopes: 204mPb, 199mHg, 111Ag Shifts: 12 (done) + 6 (2011)‏ + 7 (bio-beta-NMR) Experiments: ~75 Publications: 5 (+ 3 manuscripts in preparation) Year IS448 IS488 2009 204mPb – 4 shifts (cancelled) 199mHg – 6 shifts (50 experiments)‏ 111Ag – 3 shifts (0 experiments)‏ 2010 ------------- 199mHg – 3 shifts (23 experiments)‏ 2011 199mHg – 4 shifts (October) 111Ag – 2 shifts (July)‏ Monika Stachura INTC meeting July 2011

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

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

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

Natural proteins – HAH1 (Copenhagen & Michigan & Wroclaw)‏ A. K. Wernimont et al., Nature Structural Biology, 7, 766 - 771 (2000) Manuscript in preparation Monika Stachura INTC meeting July 2011

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

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

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