BIG GUNS National High Magnetic Field Laboratory’s Amy McKenna (left) and Rodgers get the resolution they need to dete

1. September 21, 2009 | Volume 87 | Number 38 | pp. 12 - 17 Digging Into Asphaltenes Mass spectrometry uncovers chemical details of petroleum’s most recalcitrant fraction Celia Henry Arnaud COVER STORY ACS: 158,000 Members BIG GUNS National High Magnetic Field Laboratory’s Amy McKenna (left) and Rodgers get the resolution they need to determine asphaltene composition from high-field Fourier transform ion cyclotron resonance MS.

2. 21 T FT-ICR MS ~$15M Magnet ~$2.5M Spectrometer NHMFL: NSF Chemistry Division EMSL: Dept. of Energy AMOLF: NWO Netherlands

3. Citations #1 Most-cited JASMS, 1996-2008 "External Accumulation of Ions for Enhanced Electrospray Ionization FT-ICR MS", JASMS1997, 8, 970-976 # 1 Most-cited MS Reviews, 1998-2008 "Fourier Transform Ion Cyclotron Resonance Mass Spectrometry: A Primer," Mass Spectrom. Rev. 1998, 17, 1-35 # 2 & #5 Most-cited Energy & Fuels, 2005-2008 "Reading Chemical Fine Print: Resolution and Identification of 3000 Nitrogen-Containing Aromatic Compounds from a Single Electrospray Ionization FT-ICR Mass Spectrum of Heavy Petroleum Crude Oil," Energy & Fuels, 2001, 15, 492-498. "Resolution and Identification of Elemental Compositions for More than 3000 Crude Acids in Heavy Petroleum by Negative-Ion Microelectrospray High Field FT-ICR MS," Energy & Fuels, 2001, 15, 1505-1511

4. Top-Down Proteomics: Identification and Characterization of Intact Proteins Ji Eun Leea, John F. Kelliea, John C. Trana, Jeremiah D. Tiptonb, Adam C. Cathermana, Haylee M. Thomasa, Dorothy R. Ahlfa, Kenneth R. Durbina, Adaikkalam Vellaichamya, Ioanna Ntaia, Alan G. Marshallb,c, and Neil L. Kellehera aIon Cyclotron Resonance Program,National High Magnetic Field Laboratory, Tallahassee, Florida 32310-4005 b Dept. of Chemistry and Biochemistry, Florida State Univ., Tallahassee, FL 32306 cDept. of Chemistry, U. Illinois, Urbana, Ill. 61801 The most common ("Bottom-Up") way to identify proteins is to break them into fragments with enzymes in solution, and then analyze the pieces by liquid chromatography/mass spectrometry. However, if the proteins can be ionized intact in the gas phase, and then fragmented ("Top-Down" proteomics), identification is more direct and certain, and the location(s) of chemical modifications (e.g., phosphates, sugars, etc.) become unequivocal. Recently, it has become possible to fractionate a protein mixture by gel electrophoresis, and then sort the contents by nanocapillary liquid chromatography and ultrahigh-resolution mass spectrometry and MS/MS. A single injection yields ~40 detectable proteins, about half which yield ProSight (automated) identifications. For example, the Figure shows differences in phosphorylation for proteins identified in asynchronous and M phase arrested HeLa cells. Asynchronous (A) and M-phase arrested (B) HeLa proteins are confidently identified, with the phosphorylations designated in red. A fragment map (C) is shown for the doubly phosphorylated 60S acidic ribosomal protein. At higher magnetic field (21 T), such identifications should extend to the larger proteins found in humans. *Suuported by NSF Division of Materials Research through DMR-06-54118, and the State of Florida. Lee, J. E.; Kellie, J. F.; Tran, J. C.; Tipton, J. D.; Catherman, A. C.; Thomas, H. M.; Ahlf, D. R.; Durgin, K. R.; Vellaichamy, A.; Ntai, I.; Hendrickson. C. L.; Emmett, M. R.; Marshall, A. G.; Kelleher, N. L. "A Robust 2-Dimensional Separation for Top-Down Tandem Mass Spectrometry of the Low Mass Proteome," J. Am. Soc. Mass Spectrom. 2009, 20, 0000-0000.

5. DOes a Truncated Domain Accurately Represent an Intact Enzyme: KIT Tyrosine Kinase Hui-Min Zhang,a Mark R. Emmett, a,b Michael Greig,c and Alan G. Marshalla,b,* aIon Cyclotron Resonance Program,National High Magnetic Field Laboratory, Tallahassee, Florida 32310-4005, USA b Dept. of Chemistry and Biochemistry, Florida State Univ., Tallahassee, FL 32306, USA cGlobal Research & Development, Pfizer Inc. La Jolla Laboratories, 10777, Science Center Drive, San Diego, CA  92121 The most detailed probes of drug receptor architecture are nuclear magnetic resonance and x-ray crystallography. However, it is often necessary to "snip" away all but the putative critical core "domain", to make it amenable to NMR or x-ray analysis. One then needs to validate that approach, by somehow comparing the domain and intact enzyme, to see if the domain does in fact retain the same critical structure (e.g., catalytic site and regulatory site) as the intact protein. We recently used hydrogen/deuterium exchange for such a validation for tyrosine kinase (KIT), the target for drugs (imatinib and sunitinib) to treat abdominal cancer. The Figure shows that deuterium uptake (a measure of exposure to solvent) is essentially the same for the critical segments of the enzyme, with or without the 60 amino acid segment known as the "kinetic insertion domain" (KID), thereby confirming (in this case) that the domain does accurately represent the receptor structure. *Suuported by NSF Division of Materials Research through DMR-06-54118, NIH 78359, and the State of Florida. Gajiwala, K. S.; Wu, J. C.; Christensen, J.; Deshmukh, G. D.; Diehl, W.; DiNitto, J. P.; English, J. M.; Greig, M.; He, Y.-A.; Jacques, S. L.; Lunney, E. A.; McTigue, M; Molina, D.; Quenzer, T. A.; Wells, P. A.; Yu, X.; Zhang, Y.; Zou, A.; Emmett, M. R.; Marshall, A. G.; Zhang, H.-M.; Demetri, G. "KIT Kinase Mutants Show Unique Mechanisms of Drug Resistance to Imatinib and Sunitinib in Gastrointestinal Stromal Tumor Patients," Proc. Natl. Acad. Sci. U.S.A. 2009, 106, 1542-1547.

6. O2 Class 6 5 4 DBE 3 Relative Abundance (% total) 2 1 0 15 30 35 20 40 25 Carbon Number Mmilili M. Mapolelo,a Lateefah A. Stanford,b Ryan P. Rodgers, a,b Andrew T. Yen,c Justin D. Debord,c Sam Asomaning,c and Alan G. Marshalla,b,* Chemistry of Petroleum Crude Oil Deposits: Sodium Naphthenates aIon Cyclotron Resonance Program,National High Magnetic Field Laboratory, Tallahassee, Florida 32310-4005, USA b Dept. of Chemistry and Biochemistry, Florida State Univ., Tallahassee, FL 32306, USA cBaker Hughes, 12645 W. Airport Blvd, Sugar Land, Texas 77478 Calcium and sodium naphthenates are solid deposits and emulsions formed by the interaction of naphthenic acids in crude oil with divalent (Ca2+, Mg2+) or monovalent (Na+, K+) ions in produced waters. We have determined detailed chemical compositions of calcium and sodium naphthenates from the field based on high resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). In all cases, calcium naphthenate deposits consist predominately of tetraprotic acids with a C80 hydrocarbon skeleton whereas sodium naphthenate emulsions consist mainly of specific monoprotic saturated carboxylic acids. The Figure is an abundance-weighted plot of double bond equivalents (rings plus double bonds to carbon) versus number of carbons in those sodium naphthenate deposit molecules that contain carbons, hydrogens, and two oxygen atoms. Thus, sodium naphthenates are simply saturated (i.e., (-CH2-)n carbon chains terminated in a -CO2H group containing a single C=O double bond. Understanding the specific chemical nature of such deposits is the first step toward preventing their formation. *Suuported by NSF Division of Materials Research through DMR-06-54118, and the State of Florida. Mapolelo, M. M.; Stanford, L. A.; Rodgers, R. P.; Yen, A. T.; Debord, J. D.; Asomaning, S.; Marshall, A. G. "Chemical Speciation of Calcium and Sodium Naphthenate Deposits by Electrospray Ionization FT-ICR Mass Spectrometry," Energy & Fuels 2009, 23, 349-355.

7. Factor(s) of 2 Incremental Becomes Transformative

8. 861.68 861.70 861.72 861.74 861.76 Magnitude Mode m/Δm50% = 119,242 C58H91O313C2 m/z Bitumen ESI 9.4T FT-ICR MS C57H97O3S1 Absorption Mode C60H93O3 m/Δm50% = 508,320 861.68 861.70 861.72 861.74 861.76

9. Management Access Dissemination Training Diversity

10. Management: NHMFL ICR Program Staff P.I. and Director (F.S.U.) Alan Marshall U. Florida John Eyler Sch/Sci Instrumention Chris Hendrickson Sch/Sci Biological Mark Emmett Sch/Sci Oil/Environmental Ryan Rodgers Sch/Sci Data Systems Greg Blakney Sch/Sci Informatics (t.b.a.) Sch/Sci User Program (t.b.a.) Machinist Daniel McIntosh Technician John Quinn Consultant Steve Beu Consultant Eugene Nikolaev Admin Asst Colleen Davis (Scholar/Scientists are former postdocs)

11. Management External Advisory Panel Jon Amster (U. Georgia) John Eyler (U. Florida) Mike Greig (Pfizer) Kristina Håkansson (U. Michigan) David Muddiman (N. C. State)

12. Management Access Dissemination Training Diversity

13. Access

14. Access Automated H/D Exchange Data Acquisition

15. Access Spares In Stock: <5% "Down" Time Flexible Visit Duration (1 Day to 1 Year)

16. Management Access Dissemination Training Diversity

17. Dissemination Keck Magnet 25 T (1066 MHz 1H NMR), Two-Inch Bore, Low-ppm Spatial/Temporal Inhomogeneity Initiated by: $600K Proposal Organized by: Magnet Designed & Built by: Stabilization/Shimming by: Primary Applications: ICR/MS&T ICR RES/MAG NMR/OPS EMR/NMR

18. Dissemination 25 Data Systems (MIDAS, PREDATOR) Built for Other Labs

19. Dissemination NHMFL FT-ICR MS Presentations at Non-MS Meetings Forensics Petroleum/Fuels/Hydrocarbons Geochemistry Polymers Functional Genomics Pharmaceuticals Neurochemistry/Neurooncology Systems Biology

20. Dissemination NHMFL ICR International Academic Collaborations Australia Brazil Canada Denmark England Finland Germany India Israel Korea The Netherlands Norway Russia Sweden (Red: Jointly Funded Projects)

21. Dissemination 7th N. American FT MS Conference Key West, FL 18-22 April, 2009

22. Dissemination Industrial Collaborations

23. Dissemination Biomedical Collaborations

24. Dissemination 150 NHMFL ICR Facility Users New 125 Continuing Number of Users 100 75 50 25 0 2001 2002 2003 2004 2005 2006 Year

25. 350 Dissemination 300 Cumulative Users of NHMFL ICR Facility 250 Number of Users 200 150 100 50 0 2001 2002 2003 2004 2005 2006 Year

26. Management Access Dissemination Training Diversity

27. Training All Ph.D. Students and Postdocs Work with Facility Users

28. Priming the Academic Pump (1997-2009) Helen Cooper U. Birmingham (England) Michael Freitas Ohio State U. Kristina Håkansson U. Michigan Christina Hughey Chapman U. Robert Hudgins York U. (Canada) Sasa Kazazic Rudjer Bošković(Croatia) Sunghwan Kim Kyungpook Nat'l U. (Korea) Geoff Klein Christopher Newport U. Jarrod Marto Dana Farber/Harvard Matt Renfrow U. Alabama Birmingham Touradj Solouki U. Maine Guillaume van der Rest Ecole Polytechnique (France) Yury Tsybin EPFL (Switzerland) Forest White M.I.T. Ying Xiong U. Sci. Technol. (China) Fang Wang Albert Einstein Coll. Medicine

29. Priming the Industrial Pump (1997-2009) Michael Chalmers Scripps Florida Jared Drader Isis Pharmaceuticals Jochen Friedrich Bruker Jinmei Fu BristolMyerSquibb Yulin Huang Perseptive Biosystems Priyanka Juyal Nalco Guo-Zhong Li Waters Carol Nilsson Pfizer Terry Quenzer Pfizer Jeremiah Purcell Shell Global Development Michael Senko ThermoFinnigan Stone Shi Pfizer Bruce Wilcox NanoStream Zhigang Wu Medarex Zhongqi Zhang Amgen

30. Priming the Government Lab Pump (1995-2009) Do-Gyun Kim Ctr. for Disease Control Sudarslal Nair Tata Institute (India) Ljiljana Pasa-Tolic Battelle PNNL Lateefah Stanford U.S.D.E.A.

31. ICR Program Racial Diversity: (1995-2009) Reginald Little FAMU Asst. Prof. Datiska Archie FSU Undergrad Cleon Davis FAMU Undergrad Jesus Ivan Gonzalez FSU Undergrad Kali-Nicole Hodge FAMU Undergrad Verónica Iñiguez-García FSU Undergrad David Roberson FAMU Undergrad Jade Velasquez FSU Undergrad Mmilili Mapolelo FSU Ph.D. (2010) Lateefah Stanford FSU Ph.D. Marshall HBCU Talks: N. C. A&T; Howard U.

32. Diversity Dr. Christine Hughey, 2002 Ph.D. (NHMFL ICR Program) Assistant Professor of Chemistry at Chapman University 1st place in the Undergraduate Analytical Research Progam from the Society of Analytical Chemists of Pittsburgh 200: $10K Award Dr. Kristina Hakansson, 2003 Postdoc (NHMFL ICR Program) Assoc. Professor of Chemistry at U. Michigan Ann Arbor 2004: Searle Scholar Award 2005: ASMS Research Award 2005: Eli Lilly Analytical Chemistry Award 2006: NSF CAREER Award Ms. Katye Altieri, 2007 External User (NHMFL ICR Program) Ph.D. Candidate, Rutgers U. 2008 Peter B. Wagner Memorial Award for Women in Atmospheric Sciences