Transcript No Slide Title

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.
21 T FT-ICR MS
~$15M Magnet
~$2.5M Spectrometer
NHMFL: NSF Chemistry Division
EMSL: Dept. of Energy
AMOLF: NWO Netherlands
Citations
#1 Most-cited JASMS, 1996-2008
"External Accumulation of Ions for Enhanced Electrospray Ionization
FT-ICR MS", JASMS 1997, 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
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
a Ion Cyclotron Resonance Program, National High Magnetic Field Laboratory,
Tallahassee, Florida 32310-4005
b Dept. of Chemistry and Biochemistry, Florida State Univ., Tallahassee, FL 32306
c Dept. 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.
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,*
a Ion
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
c Global 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.
Deuterium Uptake for each of 53 Enzyme
Segments after 2 hours of H/D Exchange
KID
Domain
after
30 min
Exchange
Tyrosine
with
KID Domain
WTWT
KITKIT
+/- +/KID
Domain
after
8 Kinase
min
Exchange
Tyrosine Kinase without KID Domain
KIT Fragments
*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.
Chemistry of Petroleum Crude Oil Deposits:
Sodium Naphthenates
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,*
a Ion
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
c Baker Hughes, 12645 W. Airport Blvd, Sugar Land, Texas 77478
O2 Class
6
Relative Abundance (% total)
5
4
DBE
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.
3
2
1
0
15
20
25
30
35
40
Carbon Number
*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.
Factor(s) of 2
Incremental
Becomes
Transformative
Bitumen
ESI 9.4T FT-ICR MS
Magnitude Mode
m/Δm50% = 119,242
C58H91O313C2
861.68
861.70
Absorption Mode
m/Δm50% = 508,320
861.68
861.72
861.74
m/z
C57H97O3S1
861.76
C60H93O3
861.70
861.72
861.74
861.76
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