Molecular Photonics Chemical Physics Lab, chrupn@lsu.edu Department of Chemistry Louisiana State University, Baton Rouge LA 70803

 

 

Dr Rupnik                              maglab smbol    nsf     doe 

 

 

 

Name:        Dr  Kresimir Rupnik, Ph.D.                                                         Citizen: USA\

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Office:     Department of Chemistry, Louisiana State University, Baton Rouge, LA 70803

Phone:         (225) 205-1353 (work cell phone, 24/7)    (225) 578-2945 (Chemistry, LSU)

E-mail/web: chrupn@lsu.edu or rupnik@intelliom.com     http://www.intelliome.com

Field of specialization: Chemical/Molecular Physics-photonics, Physical- Analytical- Chemistr


 

 

 

 

 

Using OM PPS methods we can highly selectively identify many key details of electron-spin structures and processes in all molecular systems and in various different spectral regions from X-ray to IR and beyond. Key decision making steps in these processes are characterized by ultrafast (femto and sub-femto) time scales and therefore require highly selective spatially-temporally and objects resolved methods � such as those that we now develop in the unique environment of cells 1-3-5 at the NHMF laboratory. With well-defined precise PPS Spatio-Temporal Electro-Magnetic Processes/structures (STEMP) based on 25T field and < 15 fs laser pulses we can identify signatures of molecular processes better than 10-9 in comparison to similar non-PPS methods. Using <20fs and 25T Split-helix magnet at NHMF lab we have developed and tested OM PPS configurations (July 2014, cell 5) that can monitor previously unseen electron-spin couplings and correlations, even in soft matter systems of our interest. Earlier, we have successfully used <100fs sub-K (3He) - high magnetic flux (~10 T, cell 1) PPS instrumentation setup at the NHMF Laboratory. Indeed, OM PPS studies can significantly improve our understanding of electronic processes and structures even in the most complex processes of paramount interest in medicine:  our recent discoveries at LSU and NHMF opened a completely new insight into the various different forms of Fe-S clusters in some key enzymes.  Clear PPS signatures of electron (de)localization in active units are for the first time detected. Such active centers are known to be involved in the reduction-oxidation, ET, catalysis, biosynthesis and photosynthesis, as well as controlled synthesis. NHMFL and LSU work at higher magnetic fields resulted in new findings about the fundamental electron spin related properties which challenge earlier model predictions. For example, our recent studies of the magnetic PPS VT-MCD signatures of the key steps in the reduction processes of some nitrogenase related Fe-S clusters and the ongoing TO studies have been accepted in the scientific community with great interest. They not only challenge previous predictions and interpretations, but also provide a solid base for novel STEMP-based electron interaction models development.

MAGLABmagnet

 

 

Recent projects:

 

Probing Unusual Hidden Electron Dynamics in Molecules of Biomedical Interest Using Soft PPS Radiation Signatures (RS) in the Framework of Attomechanics Models

Summary

Unbiased RS models were originally introduced to study ultrafast, relatively higher energy events in systems of biomedical interest [1]. That work continued with the study of causal relations between electron attodynamics and the interacting (driving) radiation by measuring characteristic electron RS, that is the observed signatures of such interactions. During the last decade our Polarization Phase Selective (PPS) High Magnetic Field (HMF) instrumentation and methods developed in the framework of new integrative attomechanics modeling significantly improved [2-3]. That allowed inclusion of soft HMF PPS RS for studies in the energy region of chemical reactions in large, complex and unstable bio-molecules, �far from equilibrium�. Our main challenges are highly selective RS extractions from large sets of interfering components. Scientifically important question we address here is: can these methods probe electrons during protein synthesis and activity, or during interactions with other molecules, like radical SAM. Contrary to what has been suggested in literature, our magnetooptic studies already provide conclusive evidence about electron locations and actions in some studied enzyme reactions. However, some of our most important findings are related to the previously unseen electron dynamics and new protein configurations. Indeed, we can separate signatures from previously indistinguishable proteins that show small structural but paramount activity differences, and we can separate (symmetrical) signatures from recently discovered unusual and previously unseen electron spin-flips [4]. Such events can be hidden or �dark� due to the overlappings and interferences. We estimate that controlling window for such events starts at tens of femtoseconds or in some cases even shorter time.

 

 

Biomedicine

 

Present published models describing electron dynamics and structure are known to be incomplete and both theoretical and experimental methods are inadequate for the broader goals of our funding agencies and the emerging needs of society in areas such as materials, environment, energy , medicine, etc. . Given those challenges and (too) many failures of methods used today to address them, new science and many existing open questions desperately require fundamentally new and alternative methods. Our integrative theoretical and experimental projects are aimed at advancing research in the areas of paramount interest to both understanding electronic structure at its own spatio-temporal level as well as applications in the areas of paramount interest to biomedicine at the protein/enzyme functions level. So far the results show significant advancement in better understanding electrons attomechanics as well as in introducing new biomedical science  and discoveries. In many aspects, the last decade of  PPS RS HMF brought a very different view and much better understanding  of electrons dynamics in a broader range of investigations: from Rydberg, valence and inner electrons to the details of electrons-electromagnetic forces couplings. Among other successes in applications, newly found science presented in this work has also potentially high impact in advancing individual medicine.

 

 Previous results and ongoing work

Dr Rupnik 

 

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(A) Reversible redox titration of ΔnifB NifEN, where a Nernst curve is represented by the solid line. Blue and red points represent samples used to collect the MCD spectra in panel B. (B) MCD spectra of ΔnifB NifEN

 

HMF-PPS measurements can improve our understanding of electronic processes in enzymes even in the most complex environments of paramount interest to biophysics and medicine, Our OMPPS high field measurements

and  findings about FeS based electronic processes challenge earlier model predictions (Rupnik at all JACS, 2011-12,  Biophysics 2014). Of interest is ongoing 12fs PPS OM study that could provide inside in selective correlated electronic events.

. (A) MCD spectra of IDS-oxidized ΔnifB NifEN (red) and ΔnifH MoFe protein (green, scaled by a factor of 0.5). (B) First derivative MCD spectra of oxidized ΔnifB NifEN (red) and ΔnifH MoFe protein (green, scaled by a factor of 0.5), showing nearly identical transitions in both cases. (C) MCD spectra of reduced ΔnifB NifEN (red) and ΔnifH MoFe protein

Dr Rupnik

Dr Rupnik

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MCD magnetization of L127DFe-MoFe Av 1 protein, at ~500mK @ 410nm, a multi-component spin system  At NHMF lab, we canobtained much higher selectivity which resulted in new information about enzymatic structures and synthesis patterns.

 

MCD magnetization of Av1 DnihB MoFe: blue, at  1.63K @ 520nm (LSU), red at 1.577K @410nm  (NHMF, this work covers 10T). Pulses~100fs, 70MHz 

FSHM measurements- magnetizations of porphyrin FeCl(TPP)  shown from  -24.5 to 24.5 T

Typical UF OM  PPS- differential PPS magnetization curve - ~15fs  , 800nm centered pulse directed into electronic transitions area.  MCD curve  is shown below. Events are driven in broadband region, 80  MHz

 

Dr Rupnik

Dr Rupnik

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FSHM measurements- MCD of porphyrin FeCl(TPP)  shown from  -24.5 to 24.5 T measurements. A larger set of porphyrins is investigated for new unexpected electronic structures

 

OM PPS spectra of a Fe4S4- protein. Temperature with sapphire windows ~10K. A shift of the peak at 520->580nm is observed at VT. Results are different from low field: different view of correlated coupled electrons

Typical UF OM PPS- differential PPS structure - ~15fs  , 800nm centered pulse directed into electronic transitions area.  Magnetization curve  is shown above from 0 to 25T. Events are driven in broadband region

High OM PPS can significantly modulate temporal phase and also help  selectively resolve diferent spatio-temporal contributions. We investigate now presence of oscillatory structures below 100fs. Here some models predict new important couplings

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Dr Rupnik

 

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OUR OM HF UF PPS ~ 15fs measurements  in various different spectral regions is based on several new PPS technological solutions

Sample preparation and presentation

technology presents one of the most

significant challenges and one of the great successes of the previous OM PPS  work at NHMFL.

At sub 15fs , STEMPS measurements and characterizations require new instrumental configurations for photonic auto- and cross-correlations

Femtosecond scale signal from a PPS.PPS method provide multidimensional data: spatial, temporal frequency, and phase information-higher selectivity and better STEMP resolution.  Polarization selection is achieved with two polarizers.

 

Professional:

1986-present   Faculty-Research Scientist,  Department of Chemistry, Louisiana State University, Baton Rouge, LA

1984-85      Postdoctoral Researcher in Physical Chemistry, Department of

                    Chemistry, University of North Carolina, Chapel Hill, NC  (NSF)

Awarded Projects/Grants: Research in the last 20 years included projects supported by NSF, USDoA, NIH, DOD, DOE and other funding agencies.

-NSF(Collaborative Investigator) � (September2012-5 years) : �Development of Ultrafast Spectroscopy System for Chemistry , Materials Science and Biophysics Research and Education in the 25-T Split-Coil Helix�

-(PI) NHMF        (2019-on) Variable Magnetic-Field (VMF) Ultrafast (UF) Polarization Phase Selective (PPS) Attomechanics Studies in the Areas of Fundamental Significance to Biochemistry/Biophysics: Towards New Methods and Tools for Monitoring Electronic Processes in Biosystems (soft matter)

- (PI) NHMF -## (new 2014-2016), �Ultrafast  STEMP Polarization Phase Selective Studies�-

- (PI) NHMF -## (20012-2014), �Ultrafast Polarization Phase Selective Studies�-

- (PI) NHMF -## (2007-2012), �Polarization Phase Selective Studies�-

-Collaborative projects Dr. Brian Hales LSU (2000-2014) and Dr Markus Ribbe  UC Irvine, (NIH  GM67626  and NSF funds, BoR, EPSCOR funds) ,

-(DOE, NIH) Collaborative projects at Argonne National Laboratory, Advanced Photon Source, 2002-present.

- DOE (Co-PI), "A Physico-Chemical Study of Some Areas of Fundamental Significance  to Biophysics" (project from 1986)

-LEQSF (PI), �Radiation Signatures� (1996)

-Collaboration on science education projects: NSF �Scientific Visualization� 2007-present, NSF �Student retention project�, 2007-present.

 

 Selected Publications:

1a.      K. Rupnk, C. C. Lee, Y. Hu, M. W. Ribbe and B. J. Hales, �A VTVH MCD and EPR Spectroscopic Study of the Biosynthesis of the �Second� Nitrogenase P-Cluster�, submitted Inorganic Chemistry �, Inorganic Chemistry, 57, 4719  (2018);

1.  K Rupnik, C. Lee, J. A. Wiig, Y. Hu, M. W. Ribbe, and B. J. Hales , �Non-Enzymatic Synthesis of the P-Cluster in the Nitrogenase MoFe Protein: Evidence for the Involvement of All Ferrous Intermediates�, Biochemistry, 55 ( 2014) 1108

2. J. Dumke, A. Qureshi, S. Hadman, S. Das, K. Rupnik, B El-Zahab, D. J. Hayes, I. M. Warner, �Tumor-Targeting Hyperthermal near Infrared Nano GUMBOS�, Photochem. Photobiol. Sci. (submitted 2014).

3. K. Rupnik PI, �NHMFL PPS Studies Project Report 2010- 2013�, NHMFL, 2013

4. K. Rupnik, Y. Hu, C. C. Lee, J. A. Wiig, M. W. Ribbe and B.J. Hales, �P+ State of Nitrogenase P-Cluster exhibits Electronic Structure of a [Fe4S4]+ Cluster�, J. Am. Chem. Soc. 134 ( 2012) 13749

5. K. Rupnik, C. C. Lee, Y. Hu, M. W. Ribbe, B. J. Hales, �[4Fe4S]2+ Clusters Exhibit Ground-State Paramagnetism�, J. Am. Chem. Soc. 133 (2011) 6871.

6. K. Rupnik, Yilin Hu, Aaron W. Fray, Markus Ribbe, and Brian J. Hales, �VTVH-MCD Spectroscopic Study of  NifEN-Bound precursor�, J. Biol. Inorg. Chem., 16  (2011) 325.

7. T.  Brown, Z. LeJeune, K. Liu, S. Hardin, J-R Li, K. Rupnik, and J. C. Garno, �Automated scanning probe lithography with n-alkanethiol self assembled monolayers on Au(111): Application for teaching undergraduate laboratories� , JALA,16 (2) (2011) 112.

8. D. Shafir, Y. Mairesse, H. J. Worner, K. Rupnik, D.M. Villeneuve, P. B. Corkum and N. Dudovich, �Probing the symmetry of atomic wavefunction from the point of view of strong field driven electrons� , New Journal of Physics 12, 073032 (2010)

9. Marcia S. Cotton, Kresimir Rupnik, Robyn B. Broach, Yilin Hu, Aaron W. Fay, Markus W. Ribbe and Brian J. Hales VTVH-MCD Study of the DnifBDnifZ MoFe Protein from Azotobacter vinelandii, J.  Am. Chem.  Soc.,  131 (13) (2009) 4559.

10. K. Rupnik PI, �NHMFL PPS Studies Project Report 2008 and 2009�,NHMFL 2009

11. R. Broach, K. Rupnik, M. Ribbe, B, J, Hales, �MCD Spectroscopic Investigations of the Different Oxidation States of the DnifH apo-MoFe protein from A. vinelandii�, Biochemistry 2006, 45  15039-15048.

12. Uwe Bergmann, Wolfgang Sturhahn, Donald E. Linn, Jr.,Francis E. Jenney, Jr., Michael W. W. Adams, Kresimir Rupnik, Brian J. Hales, Ercan E. Alp, Aaron Mayse, and Stephen P. Cramer, �Observation of Fe-H/D Modes by Nuclear Resonant Vibrational Spectroscopy�,J.  Am. Chem.  Soc. 125, 4016 (2003)

13. S. P. McGlynn, P. Brint, J. D. Scott, and K. Rupnik, in "Understanding Chemical Reactivity: The Role of Rydberg States in Spectroscopy and Photochemistry", edited by C. Sandorfy, (Kluwier Academic, Norwell, MA, 1999), p. 121.

14. A. Vrancic, K. Rupnik, L. Klasinc, and S. P. McGlynn, "Time-Resolved Profiles in Modulated Polarization Spectroscopy", J. Chem. Info. Comput. Sci.  39, 68 (1999).

15. A. Vrancic, K. Rupnik, and S. P. McGlynn, �A Selective Digital Integrator for Modulated-Polarization Spectroscopy: An Evaluation using (+)-3 Methylcyclopentanone,� Rev. Sci. Instrum. 69,40(1998).

16. K. Rupnik and S. P. McGlynn, "The Simulation of an Unusual Magnetic Circular Dichroism Spectrum: The 5p 6s Transition of HI", J. Chem. Phys. 103, 7661 (1995).

17. K. Rupnik, U. Asaf, and S. P. McGlynn, in "Gaseous Dielectrics VII" , edited by L. C. Christophorou, (Plenum, New York, 1994), p. 87.

18. K. Rupnik, L. Klasinc, M. Varma, J. Battista, and S. P. McGlynn, "Lesion Spectra: Radiation Signatures and Biological Gateways", J. Chem. Info. Comput. Sci. 34, 1054 (1994).

19. U. Asaf, K. Rupnik, G. Reisfeld, and S. P. McGlynn, "Pressure Shifts and Electron Scattering Lengths in Atomic and Molecular Gases", J. Chem. Phys. 99, 2560 (1993).

20. M. Eckert-Maksic, Z. B. Maksic, M. Hodoscek, and K. Rupnik, "Inter and Extramolecular Electrostatic Potentials in Vitamin C", J. Mol. Struct. THEOCHEM, 256, 271 (1992).

21. W. S. Felps, K. Rupnik, and S. P. McGlynn, "Electronic Spectroscopy of Cyanogen Halides", J. Phys. Chem. 95, 639 (1991).

22. K. Rupnik, U. Asaf, and S. P. McGlynn, "Electron Scattering in Dense Atomic and Molecular gases: An Empirical Correlation of Polarizability and Electron Scattering Length", J. Chem. Phys. 92, 2303 (1990).

23. K. Rupnik, "VUV and Laser Raman Study of the Atomic and Molecular Polarizabilities", in Lasers98, edited by D. G. Harris, (STS Press, McLean, VA,1990), p. 774.

24. R. G. Parr, K. Rupnik and S. Gosh, "Phase-Space Approach to the Density Functional Calculation of Compton Profiles of Atoms and Molecules", Phys. Rev. Letts.  56, 1555 (1986).