The 3rd BK21 Plus Workshop (2014) on Nanobiomaterials and Advanced Analytical Techniques
제 3 회 BK21 플러스 워크숍(2014): 나노·바이오물질과 첨단분석기술
 Dates
: Friday, November 28, 2014
 Venue : Seminar Room 31402, College of Natural Science, Changwon National
University, Changwon, South Korea
 Organization: The BK21 Plus Research Training Group for Nano-biomaterials
and Advanced Analytical Techniques
 Sponsored by: National Research Foundation of Korea (NRF)
Welcome Address
Ladies and Gentlemen:
It is a great pleasure and privilege for me to welcome all of you and chair to “The
3rd BK21 Plus Workshop (2014) on Nano-biomaterials and Advanced Analytical
Techniques” organized by the BK21 Plus Research Training Group for Nanobiomaterials and Advanced Analytical Techniques. I take this opportunity to extend to all
of you warm greetings on behalf of the organizing committee. Special thanks are due to
you all for gathering here to contribute to this workshop.
I am happy to see the positive response which we have received from scientists
working on nano-biomaterials in the bioscience. Considering the importance of this
research area for many research groups in our country and the need for domestic and
international cooperation on problems of common interest, we have decided to hold this
workshop in order to elaborate on issues relating to the development of nano-biomaterials
in the bioscience. While appreciating your response to our invitation, I hope this meeting
will prove to be productive and beneficial for the entire participants.
Today’s workshop would include numbers of advanced topics regarding the cell
membrane and glycoproteins in bioscience trials and their assessments to ensure
qualitative analysis by mass spectrometry and magnetic resonance (MR) research for
studying protein-ligand interactions. This workshop has been designed to be practical
with lectures, examples and exercises to provide skills, encourage participation, and
exchange information.
Today’s workshop is attended by 6 speakers, especially including China and
Japan, representing leading research groups in bioscience and analytical science.
Therefore, this workshop will provide us not only essential knowledge but also a great
opportunity to share technical experiences and issues.
I would like to take this opportunity to express my sincere thanks to the
organizers and in particular our honorable speakers. Finally, this is an opportune time for
me to declare the official opening of the “The 3rd BK21 Plus Workshop (2014) on Nanobiomaterials and Advanced Analytical Techniques” and I wish fruitful day of interesting
and beneficial program and also that you have a pleasant stay in Changwon.
I warmly welcome you all again.
November 28, 2014
Yong-Ill Lee
Dean of College of Natural Science
Director of the BK21 Plus Research Training Group for Nano-biomaterials and
Advanced Analytical Techniques
Changwon National University
Organization
Director of the BK21 Plus Research Training Group for Nano-biomaterials and Advanced
Analytical Techniques
Dr. Yong-Ill Lee (Department of Chemistry, Changwon National University)
Organizing Committee:
Dr. Dong-Soo Shin (Department of Chemistry, Changwon National University)
Dr. Yong-Ill Lee (Department of Chemistry, Changwon National University)
Dr. Jae-Min Lim (Department of Chemistry, Changwon National University)
Executive Organizer:
Dr. Jae-Min Lim (Department of Chemistry, Changwon National University)
Sponsors:
BK21 Plus, National Research Foundation of Korea (NRF)
PROGRAM
Opening Session
14:0014:10
Welcome Address
Yong-Ill Lee
Dean of College of Natural Science
Director of the BK21 Plus Research Training Group for Nano-biomaterials and
Advanced Analytical Techniques
Changwon National University
Session I: Recent Technologies in Biochemistry
(Chair: Jae-Min Lim, Changwon National University)
14:1014:40
[Invited Speaker_1] "Membrane Micro-domain to Membrane Nanodomain: a Paradigm Shift for Cell Biology"
Intaek Lee
Visiting Scientist, Department of Cell Biology, Yale University School of Medicine, USA
Co-Principal Investigator (with James E. Rothman), Shanghai Institute for Advanced
Immunochemical Studies, ShanghaiTech University, Shanghai, China
14:4015:10
[Invited Speaker_2] “Aberrant Glycosylation in Cancer and
Applications to Cancer Biomarker Development”
Yong-Sam Kim
Aging Intervention Research Center, Korea Research Institute of Bioscience &
Biotechnology
15:1015:40
[Invited Speaker_3] “Glycoproteome Profiling of Human Gastric
Cancer Tissues using Filter Aided Capture and Elution (FACE)
Method and LC-MS/MS”
HooKeun Lee
College of Pharmacy, Gachon University
15:4015:50
Coffee Break
Session II: Advanced Analytical Technologies
(Chair: Intaek Lee, ShanghaiTech University, China)
15:5016:20 [Invited Speaker_4] “How to Make a Movie with Mass Spectrometry?
Time-Resolved Studies on Peptide Photodissociation & Biomolecular
Trunover”
Tae-Young Kim
Division of Liberal Arts & Sciences, Gwangju Institute of Science and Technology
16:2016:50
[Invited
Speaker_5]
“Development
of
Highly
Sensitive
Oligosaccharide Analysis Method by Enzyme and Chemical Labeling”
Jun Zhe Min
School of Pharmaceutical Science, University of Shizuoka, Japan
College of Pharmacy, Yanbian University, China
16:5017:20
[Invited
Speaker_6]
“Real-time
Hyperpolarized
Magnetic
Resonance Spectroscopy and Imaging”
Youngbok Lee
Department of Applied Chemistry, College of Science & Technology
Hanyang University ERICA Campus
17:2017:30
Closing Remarks and Photo
List of Invited Speakers
Invited Speaker_1: Intaek Lee, Ph.D.
 Visiting Scientist, Department of Cell Biology, Yale University School of
Medicine, USA
 Co-Principal Investigator (with James E. Rothman), Shanghai Institute for
Advanced Immunochemical Studies, ShanghaiTech University, Shanghai, China
 E-mail: [email protected]
Invited Speaker_2: Yong-Sam Kim, Ph.D.
 Principal Investigator, Aging Intervention Research Center, KRIBB, Korea
 E-mail: [email protected]
Invited Speaker_3: Hookeun Lee, Ph.D.
 Professor, College of Pharmacy, Gachon University, Korea
 E-mail: [email protected]
Invited Speaker_4: Tae-Young Kim, Ph.D.
 Assistant Professor, Division of Liberal Arts & Sciences and Department of
Chemistry, Gwangju Institute of Science & Technology, Gwangju, South Korea
 E-mail: [email protected]
Invited Speaker_5: Jun Zhe Min, Ph.D.
 Associate Professor, School of Pharmaceutical Sciences, University of Shizuoka,
52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan
 Associate Professor, College of Pharmacy, Yanbian University, China.
 E-mail: [email protected]
Invited Speaker_6: Youngbok Lee, Ph.D.
 Assistant Professor, Department of Applied Chemistry, Hanyang University
55 Hanyangdaehak-ro, Sangnok-gu, Ansan, Gyeonggi-do, 426-791, Korea
 E-mail: [email protected]
ABSTRACTS
Invited Speaker_1
Membrane micro-domain to membrane nano-domain: A paradigm shift for cell
biology
Intaek Lee1,2
1
Shanghai Institute for Advanced Immunochemical Studies, Shanghaitech University,
Shanghai, China; 2Department of Cell Biology, Yale University School of Medicine, New
Haven, CT, USA
E-mail: [email protected]; [email protected]
With a recent advancement of fluorescent nanoscopy, there have been drastic
changes in the ways, by which traditional cell biological studies are carried out. For the
field of membrane trafficking, this new technical advancement provides the researchers
with the ability to study protein-protein and protein-membrane interaction, involved in
cargo sorting and secretion, at nano-meter scale. Thus, cell biologists are being
challenged to answer more advanced questions on spatial and temporal regulation of the
diverse membrane trafficking machineries within membrane nano-domains in living cells,
rather than within micro-domains in fixed cells. To meet these new demands, we are
currently using not only traditional cell biology and biochemistry, but also developing
new technical approaches to characterize the fundamental machineries of membrane
trafficking at nanometer scale. These include; (i) CRISPR/Cas9-mediated genome editing
for fluorescent/epitope tagging of endogenous gene products to avoid over-expressionderived artifact during live imaging and domain mapping studies; (ii) BioID, a recently
developed proteomics technique that allows in vivo tagging of neighboring proteins via
proximity-based biotinylation, to study spatial arrangement of diverse protein
machineries in living cells. We present here our recent results from BioID studies of
membrane trafficking machineries and describe how this new proteomics approach
complements cell biology in the new era of super-resolution nanoscopy.
References
Roux KJ, Kim DI, Raida M, Burke B. “A promiscuous biotin ligase fusion protein
identifies proximal and interacting proteins in mammalian cells.” J Cell Biol. 2012,
196(6):801-10.
Hyun-Woo Rhee, Peng Zou, Namrata D. Udeshi, Jeffrey D. Martell, Vamsi K. Mootha,
Steven A. Carr, and Alice Y. Ting, “Proteomic Mapping of Mitochondria in Living Cells
via Spatially Restricted Enzymatic Tagging” Science, 2013, 339:1328-1331.
Biographies of Invited Speaker_1
1.
Current Position
Visiting Scientist, Department of Cell Biology, Yale University School of Medicine,
USA
Co-Principal Investigator (with James E. Rothman), Shanghai Institute for Advanced
Immunochemical Studies, ShanghaiTech University, Shanghai, China
E-mail: [email protected]; [email protected]
2.
Education and Professional Background
B.A, Department of Molecular and Cell Biology, University of California, Berkeley,
CA, USA (1998)
Ph.D Department of Biochemistry, University of Georgia, Athens, GA, USA (2005);
Adviser: Michael Pierce, Ph.D
NIH postdoctoral fellow, Department of Internal Medicine, Washington University in
St.Louis, MO, USA (2005-2008); Adviser, Stuart Kornfeld, M.D
Postdoctoral associate, Department of Cell Biology, Yale University School of
Medicine, New Haven, CT, USA (2008-2011); Adviser, James E. Rothman, Ph.D
Associate Research Scientist, Department of Cell Biology, Yale University School of
Medicine, New Haven, CT (2011-2013)
Visiting Scientist, Department of Cell Biology, Yale University School of Medicine,
New Haven, CT (2014-present)
Co-Principal Investigator (with James E. Rothman), Shanghai Institute for Advanced
Immunochemical Studies, ShanghaiTech University (상하이과학기술대학교,
Shanghai, China (2014-present)
3.
Research Fields and Interests
Cell Biology, Membrane trafficking, Biochemistry
Our lab studies fundamental principles of membrane trafficking and protein secretory
pathway in mammalian cells. In particular, our current research interests are threefold; (i) the role of Septin family proteins in protein secretory pathway; (ii) Golgi-
associated palmitoyltransfereases zDHHCs family in nano-domain formation in the
Golgi; (iii) the role of Acyl-CoA Binding Protein3 (ACBD3;GCP60) in establishing
functional Rab domains in the medial-Golgi.
4.
Selected Publications
Intaek Lee*I, Neeraj Tiwari*, MyunHwa Dunlop*, Morven Graham, Xinran Liu and
James E. RothmanI “Membrane Adhesion Dictates Golgi Stacking and Cisternal
Morphology” Proceedings of the National Academy of Sciences, 2014 111:18491854 (I, co-corresponding author), Faculty of 1000
Intaek Lee*, Balraj Doray*, Jennifer Govero and Stuart Kornfeld “Binding of Cargo
Sorting Signals to AP-1 enhances its association with Arf1-GTP” Journal of Cell
Biology, 2008, 180:467-472 (*Equal Contribution), Faculty of 1000
Invited Speaker_2
Aberrant Glycosylation in Cancer and Applications to Cancer Biomarker
Development
Yong-Sam Kim
Aging Intervention Research Center, KRIBB
E-mail: [email protected]
Protein glycosylation plays a variety of roles including molecular interaction,
recognition, stability, etc. However, alteration in glycan structures, termed aberrant
glycosylation, is widely observed under diseased states including cancer. The
pathological roles of aberrant glycosylation in cancer progression have been exemplified
by TIMP-1, PTPk and PSGL-1 whose aberrant glycosylations exert to increased cancer
invasion and cancer metastasis. These functional studies prompted to mine cancerspecific aberrant glycoproteins as cancer diagnostic biomarkers. Multi-lectins-based
strategy enabled to develop a panel of HCC biomarkers whose core-fucosylation, when
normalized by total amounts, may be an indicator for cancer development. This result
was obtained by a developed validation method in which biomarker-specific antibodies
are tagged with oligonucleotides each with identifiable DNA sequence. The nucleotides
are used as a template for generation of fluorescent mRNA and the fluorescent mRNAs
are measured by DNA microarray. These approaches allowed us to develop HCC
diagnostic system with a panel of HCC glyco-biomarkers. This aberrant-glycoproteins
targeted cancer biomarker development conferred an enhanced diagnostic specificity over
existing diagnostic systems.
Biographies of Invited Speaker_2
1.
Current Position
Principal Investigator at the KRIBB
2.
Education and Professional Background
Dept. of Agricultural Chemistry, Seoul Nat’l University BS
1991-1996
Dept. of Agricultural Chemistry, Seoul Nat’l University MS
1996-1998
1998-2002
2002-2003
2003-2006
2006-2012
2011
2012-current
3.
Dept. of Agricultural Chemistry, Seoul Nat’l
PhD
University
Seoul National University
Research associate
Korea Institute of Bioscience and Postdoctoral fellow
Biotechnology
Korea Institute of Bioscience and Senior researcher
Biotechnology
FHCRC, Seattle, WA, USA
Research associate
Korea Institute
Biotechnology
of
Bioscience
and Principal Investigator
Research Fields and Interests
Cancer biology, Biomarker Discovery, Glycobiology
4.
Selected Publications
Ju Hee Lee, Chang Hee Cho, Sun Hee Kim, Jeong Gu Kang, Jong Shin Yoo,
Chulhun Ludgerus Chang, Jeong-Heon Ko, Yong-Sam Kim (2014) Quantitative
measurement of a specific glycoform using a DNA-tagged antibody and lectin
affinity chromatography for glyco-biomarker development. Mol. Cell. Proteome.
Under revision.
Yong-Sam Kim, Yeong Hee Ahn, Kyoung Jin Song, Jeong Gu Kang, Ju Hee Lee,
Seong Kook Jeon, Hyoung-Chin Kim, Jong Shin Yoo, Jeong-Heon Ko (2012)
Overexpression and β-1,6-N-acetylglucosaminylation-initiated aberrant glycosylation
of TIMP-1: a "double whammy” strategy in colon cancer progression. J. Biol. Chem.
287(39), 32467-32478.
Yeong Hee Ahn*, Yong-Sam Kim*, Eun Sun Ji, Ji Yeon Lee, Ji-Ae Jung, Jeong
Heon Ko and Jong Shin Yoo (2010) Comparative quantitation of aberrant
glycoforms by lectin-based glycoprotein enrichment coupled with multiple-reaction
monitoring mass spectrometry. Anal. Chem. 82(11), 4441-4447. * Co-first author
Yong-Sam Kim, Ok Lye Son, Ju Yeon Lee, Sun Hee Kim, Sejeong Oh, Yoon Suk
Lee, Cheorl-Ho Kim, Jong Shin Yoo, Jeong-Hwa Lee, Eiji Miyoshi, Naoyuki
Taniguchi, Samir M. Hanash, Hyang Sook Yoo, and Jeong Heon Ko. (2008) Lectin
precipitation using phytohemagglutinin-L4 coupled to avidin-agarose for serological
biomarker discovery in colorectal cancer. Proteomics 8, 3229-3235
Yong-Sam Kim, Soo Young Hwang, Hye-Yeon Kang, Hosung Sohn, Sejeong Oh,
Jin-Young Kim , Jong Shin Yoo, Young Hwan Kim, Cheorl-Ho Kim, Jae-Heung
Jeon, Jung Mi Lee, Hyun Ah Kang, Eiji Miyoshi, Naoyuki Taniguchi, Hyang-Sook
Yoo, Jeong-Heon Ko (2008) Functional proteomics study reveals that Nacetylglucosaminyltransferase V reinforces the invasive/metastatic potential of colon
cancer through aberrant glycosylation on TIMP-1. Mol. Cell. Proteom. 7, 1-14.
Hosung Sohn*, Yong-Sam Kim*, Hyun-Taek Kim, Cheol-Hee Kim, Eun-Wie Cho,
Hye-Yeon Kang, Nam-Soon Kim, Cheorl-Ho Kim, Seong Eon Ryu, Jeong-Hwa Lee,
Jeong Heon Ko (2006) Ganglioside GM3 is involved in neuronal cell death. FASEB
J. 20, 1248-1250. * Co-first author
Invited Speaker_3
Glycoproteome profiling of Human Gastric Cancer Tissues using Filter Aided
Capture and Elution (FACE) method and LC-MS/MS
HooKeun Lee
College of Pharmacy, Gachon University
E-mail: [email protected]
Protein glycosylation is the most important post-translational modifications
(PTMs). Glycoproteins are often important integral proteins that play a role in cell-cell
interactions,, developmental biology, cell signaling and inflammation in cell membrane.
Because of low stoichiometry of modified proteins, for identifying the numerous
glycoproteins, the proteomics analysis of these PTM requires powerful and indispensable
technology. To detect low abundant glyco-peptides or proteins in complex mixture
among the high abundant non glycopeptide equivalents with LC-MS technologies,
enrichment methods have to be applied. For example hydrazide bead, ERLIC
(Electrostatic Repulsion Hydrophilic Interaction Chromatography), HILIC (Hydrophilic
Interaction liquid Chromatography), lectin affinity chromatography method have widely
been used for enrich glycol- peptides or proteins. In this study, we introduce a powerful
protocol for enrichment of glycopeptides, and analyzed glycosylation of human gastric
cancer tissue using membrane based modified FACE (filter aided capture and elution)
method. So we were identified 3,000 N-glycosylated peptide. And then, we functional
studies of the normal samples compared with cancer sample using IPA (Ingenuity
Pathway Analysis) from the list of identified glycopeptides.
Biographies of Invited Speakers_3
1.
Current Position
Professor, Gachon University
2.
2014
Education and Professional Background
Associate Professor, Gachon University
2008
Senior Research Scientist, ETH Zurich, Switzerland
2004
Research Scientist, Institute for Systems Biology, Seattle, USA
2001
Postdoc, Institute for Systems Biology, Seattle, WA, USA
2000
Postdoc, School of Mines, Golden, CO, USA
1998
Ph.D. (Physical-Analytical Chemistry) University of Utah, Salt Lake City, UT, USA
1998
3.
M.S. (Analytical Chemistry), Yonsei University, Korea
1990
B.S. (Chemistry), Yonsei University, Korea
1988
Research Fields and Interests
Disease Biomarker Discovery using Quantitative Proteomic Technologies
Mass spectrometric characterization of Biologics
Clinical Lipidomics
4.
Selected Publications
Tran T. H.; Park S. Y.; Lee H.; Kim B.; Kim O.-H.; Oh B.-C.; Lee D.; Lee H. “Ultra
Small Gold Nanoparticles for highly specific Isolation/Enrichment of N-linked
Glycosylated peptides” Analyst, 2012, 137, 991-8.
Tran T.; Kim O.-K.; Park J.-M.; Kim B.; Choi D.-Y.; Lee J.; Kim K.; Oh B.-C.; Lee
H. “Combined phospho- and glycoproteome enrichment in nephrocalcinosis tissues
of phytate-fed rats” Rapid Commun. Mass Spectrom., 2013, 27, 2767-2776.
Brunner E.; Ahrens C. H.; Mohanty S.; Baetschman H.; Loevenich S.; Potthast F.;
Deutsch E. W.; Panse C.; deLichtenberg U.; Rinner O.; Lee H.; Pedrioli P. G. A.;
Malmström J.; Koehler K.; Schrimpf S.; Krijgsveld J.; Kregenow F.; Heck A. J.;
Hafen E.; Schlapbach R.; Aebersold R. “A high quality catalog of the Drosophila
melanogaster proteome” Nat. Biotechnol. 2007, 25, 576-583.
Invited Speaker_4
How to Make a Movie with Mass Spectrometry? Time-Resolved Studies on Peptide
Photodissociation & Biomolecular Trunover
Tae-Young Kim
Division of Liberal Arts & Sciences, Gwangju Institute of Science & Technology.
E-mail: [email protected]
Mass spectrometry (MS) has become a standard platform for the analysis of
biomolecules since the advent of matrix-assisted laser desorption/ionization and
electrospray ionization (ESI). The structural information needed for the identification and
characterization of biomolecules is obtained typically from tandem MS. Ultraviolet (UV)
photodissociation (PD) deposits a well-defined high energy into ions of interest in a very
short timescale and results in tandem mass spectra that are dissimilar to those observed
with other fragmentation methods. A hybrid linear ion trap/time-of-flight mass
spectrometer with an ESI source was developed for UV PD of biomolecules, particularly
peptide ions. This instrument enabled time-dependent observations of product ions
generated by 157 nm PD of peptide ions. Time-resolved detection of UV PD fragment
ions of phosphopeptides also made it possible to measure the timescale on which the
phosphate group is lost from either precursor or product ions.
Mass spectrometric measurement of biomolecular turnover is the second part of
the presentation. To study biomolecular turnover, incorporation into biomolecule of a
tracer is measured over a time. Heavy water (2H2O) has been employed as a tracer in
turnover studies. The fractional synthesis rates of cardiac mitochondrial proteins have
been measured using 2H2O labeling. The basic principles relevant to these studies and
results are presented.
References
Kim, T.-Y.*; Wang, D.*; Kim, A. K.*; Lau, E.*; Lin, A. J.; Liem, D. A.; Zhang, J.; Lam,
M. P. Y.; Zong, N. C.; Ping, P. “Metabolic Labeling Reveals Proteome Dynamics of
Mouse Mitochondria” Mol. Cell. Proteomics 2012, 11, 1586-1594.
Kim, T.-Y.; Schwartz, J. C.; Reilly, J. P. “Development of a Linear Ion Trap/OrthogonalTOF Mass Spectrometer for Time-Dependent Observation of Product Ions by Ultraviolet
Photodissociation of Peptide Ion” Anal. Chem. 2009, 81, 8809-8817.
Kim, T.-Y.; Thompson, M. S.; Reilly, J. P. “Peptide Photodissociation at 157 nm in a
Linear Ion Trap Mass Spectrometer” Rapid Commun. Mass Spectrom. 2005, 19, 16571665.
Biographies of Invited Speaker_4
1.
Current Position:
Assistant Professor, Division of Liberal Arts & Sciences, Gwangju Institute of
Science & Technology
2.
Education and Professional Background
[Education]
PhD, Analytical Chemistry (Minor: Biochemistry), Oct 2009
Indiana University, Bloomington, IN, USA
Dissertation:
Development of a Hybrid Tandem Mass Spectrometer for UV
Photodissociation of Biomolecules
Advisor: Prof. James P. Reilly
MS, Analytical Chemistry, Feb 2001
Seoul National University, Seoul, Korea
Dissertation:
Chiral Separation of 9-Fluorenylmethyl Chloroformate- and Dansyl
Chloride-Derivatized D,L-Serine by -Cyclodextrin-bonded High Performance
Liquid Chromatography
Advisor: Prof. Hie-Joon Kim
BS, Chemistry, Feb 1999
Seoul National University, Seoul, Korea
[Professional Background]
Assistant Professor, Division of Liberal Arts & Sciences, Gwangju Institute of
Science & Technology,Gwangju, Korea
Mar 2013 – Present
Postdoctoral Fellow, NHLBI Proteomics Center, David Geffen School of Medicine,
University of California, Los Angeles, CA, USA
Sep 2010 – Feb 2013
Postdoctoral Scholar, Division of Chemistry and Chemical Engineering, California
Institute of Technology, Pasadena, CA, USA
Oct 2009 – Sep 2010
3.
Research Fields and Interests
Mass Spectrometry, Proteomics, Biomolecular Turnover.
4.
Selected Publications
Kim, T.-Y.*; Wang, D.*; Kim, A. K.*; Lau, E.*; Lin, A. J.; Liem, D. A.; Zhang, J.;
Lam, M. P. Y.; Zong, N. C.; Ping, P. “Metabolic Labeling Reveals Proteome
Dynamics of Mouse Mitochondria” Mol. Cell. Proteomics 2012, 11, 1586-1594.
Kim, T.-Y.; Schwartz, J. C.; Reilly, J. P. “Development of a Linear Ion
Trap/Orthogonal-TOF Mass Spectrometer for Time-Dependent Observation of
Product Ions by Ultraviolet Photodissociation of Peptide Ion” Anal. Chem. 2009, 81,
8809-8817.
Kim, T.-Y.; Thompson, M. S.; Reilly, J. P. “Peptide Photodissociation at 157 nm in a
Linear Ion Trap Mass Spectrometer” Rapid Commun. Mass Spectrom. 2005, 19,
1657-1665.
Kim, T.-Y.; Brun, Y. V.; Reilly, J. P. “Effects of Tryptic Peptide Esterification in
MALDI Mass Spectrometry” Anal. Chem. 2005, 77, 4185-4193.
Kim, T.-Y.; Kim, H.-J. “Chiral Separation of 9-Fluorenylmethyl Chloroformate- and
Dansyl
Chloride-Derivatized
D,L-
-Cyclodextrin-Bonded
Performance Liquid Chromatography” J. Chromatogr. A 2001, 933, 99-106.
High-
Invited Speaker_5
Development of highly sensitive oligosaccharide analysis method by enzyme and
chemical labeling
Jun Zhe Min, 1Toshimasa Toyo’oka
1,2
1
School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku,
Shizuoka 422-8526, Japan; 2 College of Pharmacy, Yanbian University, China
E-mail: [email protected]; [email protected]
With Boc-Asn-GlcNAc as a basic structure, four permanently positively charged
kinds of new acceptors were synthesized as acceptors for the resolution of
oligosaccharides in glycopeptides. The synthesized acceptors enzymatically reacted with
Disialo-Asn (donor) in the presence of Endo-M. The reaction yields of each
transglycosylation product were not obvious, because we do not have all the authentic
Disialo-Asn-Bocacceptors. Therefore, we used the peak area of the transglycosylation
product detected by mass spectrometry and evaluated the utility of each acceptor. Among
the Boc-Asn-GlcNAc acceptors, the positively charged MPDPZ derivative peak area was
the highest, MPDPZ-Boc-Asn-GlcNAc with a positively charged structure showed about
a 2.2 times greater sensitivity of the transglycosylation product compared to the
conventional fluorescence acceptor DBD-PZ-Boc-Asn-GlcNAc. As a result, the MPDPZBoc-Asn-GlcNAc acceptor was suitable for the transglycosylation reaction with Endo-M.
The development of a qualitative determination method for the N-linked oligosaccharides
in glycoproteins was attempted by combination of the transglycosylation reaction and
semi-micro high-performance liquid chromatography/electrospray ionization quadrupole
time-of-flight tandem mass spectrometry (HPLC/ESIQTOF-MS/MS). The asparaginyloligosaccharides in glycoproteins, liberated by treatment with Pronase E, were separated,
purified and labeled with positively charged MPDPZ. The resulting derivatives were
separated by a semi-micro HPLC system. The eluted N-linked oligosaccharide derivatives
were then introduced into a QTOF-MS instrument and sensitively detected in the ESI+
mode. Various fragment ions based on the carbohydrate units appeared in the
MS/MSspectra. Among the peaks, m/z 782.37 corresponding to MPDPZ-Boc-AsnGlcNAc is the most important one for identifying the asparaginyl-oligosaccharides.
Disialo-Asn-Boc-MPDPZ was easily identified by the selected-ion chromatogram at m/z
782.37 by MS/MS detection. Therefore, the identification of N-linked oligosaccharides in
glycoproteins seems to be possible by the proposed semi-micro HPLC separations
followed by the QTOF-MS/MS detection.
References
J. Z. Min, Y. Suzuki, Y. Tomiyasu, D. Jin, T. Higashi, Y-I Lee and T. Toyo'oka,
“Development of novel active acceptors possessing a positively charged structure for the
transglycosylation reaction with Endo-M and their application to oligosaccharides
analysis.” Rapid Commun. Mass Spectrom., 2011, 25, 2911-2922.
T. Kurihara, J. Z. Min, T. Toyo'oka, T. Fukushima and S. Inagaki, “Determination of
fluorescence-labeled asparaginyl-oligosaccharide in glycoprotein by reversed-phase ultraperformance liquid chromatography with electrospray ionization time-of-flight mass
spectrometry” Anal. Chem. 2007, 79, 8694-8698.
Biographies of Invited Speaker_5
1.
Current Position
School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku,
Shizuoka 422-8526, Japan; College of Pharmacy, Yanbian University, China.
2.
Education and Professional Background
B.S, Faculty of Pharmaceutical Sciences, Yanbian University, China (1999)
M.S, Graduate School of Pharmaceutical Sciences, University of Shizuoka, Japan
(2004)
Ph.D Graduate School of Pharmaceutical Sciences, University of Shizuoka, Japan
(2007)
Investigator Yanbian University Herbage Pharmaceutical Co., Ltd., China (19992000)
Associate Professor, School of Pharmaceutical Sciences, University of Shizuoka,
Japan (2007-present)
Associate Professor, College of Pharmacy, Yanbian University, China
(2013-
present)
3.
Research Fields and Interests
Metabonomics, Glycomics, Bioanalytical Chemistry
(1) Rapid and sensitive analysis of designer drugs.
(2) Explorations of the disease markers of human nail or saliva and the utility of the
clinical analysis sample.
(3) Development of oligosacchride highly sensitive analysis method by enzyme and
chemical fluorescent labeling.
(4) Research on the physiological significance of D-amino acid in mammals.
(5) Development of new mass derivatization reagents for resolution of chiral amines.
4.
Selected Publications
J. Z. Min, A. Matsumoto, G. Li, Y-Z Jiang, H. Yu, K. Todoroki, K. Inoue and T.
Toyo'oka, “A quantitative analysis of the polyamine in lung cancer patient
fingernails by LC-ESI-MS/MS” Biomed. Chromatogr., 2014, 28, 492-499.
J. Z. Min, M. Yamamoto, H. Yu, T. Higashi and T. Toyo'oka, “Rapid and sensitive
determination of the intermediates of advanced glycation end products in human nail
by UPLC-ESI-TOF-MS” Anal. Biochem., 2012, 424, 187-194.
J. Z. Min, Y. Suzuki, Y. Tomiyasu, D. Jin, T. Higashi, Y-I Lee and T. Toyo'oka,
“Development of novel active acceptors possessing a positively charged structure for
the
transglycosylation
reaction
with
Endo-M
and
their
application
to
oligosaccharides analysis” Rapid Commun. Mass Spectrom., 2011, 25, 2911-2922.
J. Z. Min, S. Hatanaka, T. Toyo'oka, S. Inagaki, R. Kikura-Hanajiri, Y. Goda, “Rapid,
sensitive and simultaneous determination of fluorescence-labeled designated
substances, controlled by the Pharmaceutical Affairs Law in Japan, by ultraperformance liquid chromatography coupled with electrospray-ionization time-offlight mass spectrometry” Anal. Bioanal. Chem., 2009, 395: 1411-1422.
J. Z. Min, T. Toyo’oka, T. Kurihara, T. Fukushima, S. Inagaki, “Fully automated
two-dimensional high-performance liquid chromatography with electrospray
ionization time-of-flight mass spectrometry for the determination of oligosaccharides
in glycopeptides after enzymic fluorescence labeling” J. Chromatogr. A., 2007, 1160
120-127.
Invited Speaker_6
Real-time Hyperpolarized Magnetic Resonance Spectroscopy and Imaging
Youngbok Lee
Department of Applied Chemistry, Hanyang University
E-mail: [email protected]
Magnetic resonance (MR) research is one of the most important analytical tools
for chemistry and biological study. It provides not only detailed information on the
structure of small molecules and macromolecules, but also on molecular interactions.
Because of the inherent low sensitivity of MR, which stems from a small Zeeman
splitting of the nuclear spin energy states, a long signal averaging time or a high spin
concentration is often required. A variety of methods have been explored to improve the
sensitivity of MR. Especially, large signal gains can be obtained by hyperpolarization of
the nuclear spins. MR signals of hyperpolarized samples are enhanced by several orders
of magnitude when compared to the signals from thermal polarization. Dissolution
Dynamic Nuclear Polarization (D-DNP) is a versatile technique capable of polarizing
many different nuclei in the solid state at low temperature, and subsequently providing a
hyperpolarized liquid sample following a dissolution step. The resulting signal
enhancement has made it possible to obtain detailed information in research fields as
varied as metabolic imaging or enzyme catalysis. This research aims to extend the
applicability of dissolution DNP into new areas of chemistry and biology, which involve
studying protein-ligand interactions, real-time kinetic and mechanistic studies for enzyme
catalyzed and polymerization reactions, and 13C metabolic imaging for early-stage cancer
detection.
References
J. H. Ardenkjaer-Larsen, B. Fridlund, A. Gram, G. Hansson, L. Hansson, M. H. Lerche, R.
Servin, M. Thaning and K. Golman, Proc. Natl. Acad. Sci. USA, 2003, 100, 10158-10163.
F. A. Gallagher, M. I. Kettunen, S. E. Day, D. E. Hu, J. H. Ardenkjaer-Larsen, R. Zandt, P.
R. Jensen, M. Karlsson, K. Golman, M. H. Lerche and K. M. Brindle, Nature, 2008, 453,
940-943.
Biographies of Invited Speaker_6
1.
Current Position
Assistant Professor, Department of Applied Chemistry, Hanyang University
2.
Education and Professional Background
(1) Education
Ph. D., Department of Chemistry, Texas A&M University, College Station, TX 2013
M. S., Department of Chemistry, Hanyang University, Seoul, Korea
2006
B. S., Department of Chemistry, Hanyang University, Seoul, Korea
2004
(2) Professional Career
Assistant Professor, Department of Applied Chemistry, Hanyang University
2014 - present
Postdoctoral Fellow, University of Texas MD Anderson Cancer Center 2013-2014
Visiting Researcher, University of Iowa
3.
2004-2005
Research Fields and Interests
Hyperpolarized Magnetic Resonance Imaging and Spectroscopy
NMR based Metabolomics
Early-stage Cancer Diagnosis
Real-time Kinetic and Mechanistic Study for Chemical/Biochemical Reactions
4.
Selected Publications
Lee, Y.; Zacharias, N.M.; Piwnica-Worms, D.; Bhattacharya, P.K. “Chemical
reaction-induced multi-molecular polarization (CRIMP),” Chemical Communication
2014, 50, 13030-13033.
Lee, Y.; Heo, G.S.; Zeng, H.; Wooley, K.; Hilty, C.B. “Detection of living anionic
species in polymerization reaction using hyperpolarized NMR,” Journal of the
American Chemical Society 2013, 135, 4636–4639.
Lee, Y.; Zeng, H.; Ruedisser, S.; Gossert, A.D.; Hilty, C.B. “Nuclear magnetic
resonance of hyperpolarized fluorine for detection of protein-ligand interactions,”
Journal of the American Chemical Society 2012, 134, 17448–17451. Highlighted in
Nature SciBX.
Lee, Y.; Zeng, H.; Mazur, A.; Wegstroth, M.; Carlomagno, T.; Reese, M.; Lee, D.;
Becker, S.; Griesinger, C.; Hilty, C.B. “Hyperpolarized binding pocket NOE for
determination of competitive ligand binding,” Angewandte Chemie International
Edition 2012, 51, 5179–5182. Highlighted in BRIC.
Campus Map
1. http://www.changwon.ac.kr/
2. http://w3.changwon.ac.kr/kor/campus_tour/campus_tour.html
Memorandum
Memorandum
Memorandum