Our research focuses on using mass spectrometry (MS) to understand
important biological problems, and we are particularly interested
in DNA damage induced by reactive oxygen species and covalent
modifications of proteins.
Oxidative DNA Damage
this research area, we employ a plethora of approaches,
which include analytical chemistry, synthetic organic
chemistry, biochemistry, and molecular biology, to
understand the biological implications of oxidative DNA
damage at the molecular level. Our emphasis has been placed
on the identification, structure elucidation, and
quantification of a class of oxidative DNA lesions where two
neighboring nucleobases in the same strand are covalently
linked. In this respect, we have been able to characterize
the structures of a number of novel oxidative intrastrand
crosslink lesions by mass spectrometric and NMR
In addition, we quantify the amounts of DNA lesions formed
from a variety of oxidation conditions by LC-MS/MS with
Moreover, we characterize the biophysical and biochemical
properties of oxidative DNA Lesions, and we are
particularly interested in investigating the thermodynamic
as well as the in vitro and in vivo mutagenic
properties of various oxidative DNA lesions.
2. Covalent Modifications of Proteins
projects in this area encompass the post-translational
modifications of chromosomal high-mobility group (HMG)
proteins, histones, DNA repair enzymes, and methyl-CpG
binding proteins. In
this respect, we use mass spectrometry for the
identification of the sites and types of modifications, and
we also examine the biological functions of these
have in our laboratory a ThermoFinnigan Deca XP ion trap and
an IonSpec FT-ICR mass spectrometer. We also have access to
an Applied Biosystems Voyager DE-STR MALDI-TOF and a
Micromass Global Ultima Q-TOF mass spectrometers in the
Analytical Chemistry Instrumentation Facility as well as a
Q-STAR XL O-MALDI Q-TOF mass spectrometer in the W. M. Keck
Proteomics Laboratory. Students in our lab have the
opportunity to gain hands-on experience with various
state-of-the-art mass spectrometers. Depending on the nature
of individual research project, they are also exposed to
synthetic organic chemistry, biochemistry, molecular
biology, and molecular modeling.
group picture taken in front of the new chemistry
building (Physical Sciences I, occupied in
HPLC isolation and mass spectrometric characterization of two
isomers of thymine glycols in oligodeoxynucleotides. Chem.
Res. Toxicol., 2002, 15, 671-676.
Y.*, Liu, Z., and Dixon, C. Major adenine products from
2-methyl-1,4-naphthoquinone-sensitized photoirradiation at
Biophys. Res. Commun. 2002, 291, 1252-1257.
Y.*, Vivekananda, S, and Zhang, K. ESI-MS/MS for the
differentiation of diastereomeric pyrimidine glycols in
2002, 74, 4505-4512.
Y.*, Men, L., and Vivekananda, S. Fragmentation of
deprotonated ions of oligodeoxynucleotides carrying a
5-formyluracil or 2-aminoimidazolone. J. Am. Soc. Mass
Spectrom. 2002, 13, 1190-1194.
Y.* and Liu, Z.
Mechanisms for the formation of major oxidation products of adenine
upon 365-nm irradiation with 2-methyl-1,4-naphthoquinone as
J. Org. Chem., 2002, 67, 8507-8512.
Z., Gao, Y. and Wang, Y.* Identification and
characterization of a novel crosslink lesion in d(CpC) upon
365 nm-irradiation in the presence of
Acids Res. 2003,
Q. and Wang, Y.* Independent generation of
5-(2’-deoxycytidinyl)methyl radical and the formation of a
novel crosslink lesion between 5-methylcytosine and guanine.
Am. Chem. Soc. 2003,
Y. and Wang, Y.*
Structure elucidation of DNA interstrand crosslink lesions by a
combination of nuclease P1 digestion with mass spectrometry.
2003, 75, 6306-13.
Y.*, Vivekananda, S., Men, L., and Zhang, Q.
of protonated ions of peptides containing cysteine, cysteine
sulfinic acid, and cysteine sulfonic acid.
J. Am. Soc. Mass Spectrom. 2004,
Y.; Jiang, X.; Wang, Y.*
of novel in
vivo phosphorylation sites in high mobility group N1
protein from the MCF-7 human breast cancer cells.
2004, 43, 6322-9.
C. and Wang, Y.* LC-MS/MS identification and yeast polymerase h
bypass of a novel g-irradiation-induced
intrastrand crosslink lesion G[8-5]C. Biochemistry, 2004,
Y. and Wang, Y.*
Facile formation of an intrastrand crosslink lesion between
cytosine and guanine upon Pyrex-filtered UV light
irradiation of d(BrCG) and duplex DNA containing
Am. Chem. Soc.
Y., Zhang, Q. and Wang, Y.*
mass spectrometry for the determination of the sites of DNA
Am. Soc. Mass Spectrom.
2004, 15, 1565-71.
Q. and Wang, Y.* Independent generation of the
5-hydroxy-5,6-dihydrothymidin-6-yl radical and its
reactivity in dinucleoside monophosphates. J. Am. Chem.
Soc. 2004, 126, 13287-97.
Z.; Gao, Y.; Zeng, Y.; Fang, F.; Chi, D.; Wang, Y.*
and characterization of a novel crosslink lesion in d(CpC)
induced by one-electron photooxidation.
Photochem. Photobiol. 2004, 80, 209-15.
X. and Wang, Y.* b-elimination combined with tandem mass spectrometry for the
identification of the in vivo and in vitro
sites of phosphorylation of dehydrin DHN1 protein. Biochemistry,
2004, 43, 15567-15576.
L. and Wang, Y.*
Further studies on the fragmentation of protonated ions of peptides
containing aspartic acid, glutamic acid, cysteine sulfinic
acid, and cysteine sulfonic acid.
Commun. Mass Spectrom.,
2005, 19, 23-30.
Q. and Wang, Y.* Generation of 5-(2’-deoxycytidinyl)methyl
radical and the formation of intrastrand cross-link lesions
2005, 33, 1593-1603.
Y. and Wang, Y.* Tandem mass spectrometry for the
examination of the post-translational modifications of
high-mobility group A1 proteins:
and asymmetric dimethylation of Arg25 in HMGA1a protein.
Biochemistry, 2005, 44, 6293-6301.
C. and Wang, Y.* Thermodynamic and in-vitro replication
studies of an intrastrand crosslink lesion G[8-5]C.
H. and Wang, Y.* Formation
of intrastrand crosslink products between cytosine and
adenine from UV irradiation of d(BrCA) and duplex
DNA containing a 5-bromocytosine.
Am. Chem. Soc.,
Wang, Y.* The reactivity of the
5-hydroxy-5,6-dihydrothymidin-6-yl radical in
Chem. Res. Toxicol.,
2005, in press.