Tuesday August 19, 2003
6:00 pm to 10:00 pm
Basque Cultural Center
South San Francisco, CA
Dinner reservations required
by noon on Friday August 15, 2003
(click here
for late registration information)
Quantitative Determination of Noncovalent Binding Interactions Using Automated Nanoelectrospray Mass Spectrometry
Colleen Van Pelt and Sheng Zhang
Advion BioSciences, Inc.
Ithaca, NY
Abstract
Electrospray ionization mass spectrometry (ESI/MS) has proven to be
an extremely powerful tool for studying biomolecular structures and noncovalent
interactions. Recently we reported (1) a method using a fully automated,
chip-based nanoESI/MS system to determine the dissociation constants (Kd)
for the complexes of two different proteins with their ligands. The
automated nanoelectrospray system, consisting of the NanoMate and ESI chip,
serves functionally as a combination of autosampler and nanoelectrospray
ionization source. This system provides all the advantages of conventional
nanoelectrospray plus automated, high throughput analyses without carryover.
The automated nanoESI system was used to investigate quantitative noncovalent
interactions between ribonuclease A (RNase A) and cytidic acid ligands
(2'-CMP and CTP), a well-characterized model protein-ligand complex, and
between an inactive endocellulase mutant (Thermobifida fusca Cel6A D117Acd)
and four oligosaccharide ligands (cellotriose, cellotetraose, cellopentaose
and cellohexaose). Both titration and competitive binding approaches were
performed prior to automated nanoESI/MS analysis with a Q-TOF mass spectrometer.
Dissociation constants for each complex were calculated from the sum of
ion peak areas of free and complexed proteins during the titration and
competition experiments. The measured Kd values for the RNase A-CMP
and Cel6A D117Acd-G3 complexes were found to be in excellent agreement
with the available published values obtained by standard spectroscopic
titration techniques. To our knowledge, this is the first report of using
an ESI/MS approach to study the interactions between a cellulase and oligosaccharides.
The results provide new insights for understanding the nature of cellulase-cellulose
interactions.
1. Colleen Van Pelt, Sheng Zhang and David B. Wilson, “Quantitative
Determination of Noncovalent Binding Interactions Using Automated Nanoelectrospray
Mass Spectrometry”, Anal. Chem., 2003, 75, (13) 3010-3018.
Background
Colleen Van Pelt joined Advion BioSciences as a post-doctorate student
in 1999, and has 5 years experience in nanoelectrospray mass spectrometry
with 15 publications and 2 patents. She earned her undergraduate
degree in Chemistry and French from Hartwick College. Her doctorate
was conducted at Cornell University focusing on gas-phase ion-molecule
reactions using an ion trap mass spectrometer. Dr. Van Pelt is currently
the Manager of Applications Development.
Meeting details
| Date: | Tuesday | August 19, 2003 |
| Time: | 6:00 pm | Social hour, registration (no-host cocktails) |
| 7:00 pm | Dinner | |
| 8:00 pm | Lecture | |
| Dinner: | Choice of: | (tbd) |
| includes | (various side dishes & dessert) | |
| Cost: | $25.00 | BAMS members. Reservations required by noon on Friday August 15, 2003 |
| $35.00 | Non-members. Reservations required by noon on Friday August 15, 2003 | |
| $15.00 | Students only. Reservations required by noon on Friday August 15, 2003 |
Note: 2003 dues need to be paid to obtain member price. Dues ($20) may be paid while registering for dinner.
Maps & directions
The Basque Cultural Center
599 Railroad Avenue
South San Francisco, CA 94080
Yahoo
map
Next meeting:
September 16, 2003
Recent Advances in High Resolution Microscale LC/MS for
Proteomic Analysis
Dr. Barry Karger
Northeastern University
Joint meeting with CaSSS
click here
for more information
20th Montreux LC/MS meeting, short courses and exhibition
Oct 13-17, 2003 Savannah, GA, USA
Deadlines for abstract submission and advanced registration for The
Montreux
LC/MS Symposium, to be held Oct 15-17/2003 in Savannah, GA, are approaching.
- September 1 is the deadline for submitting abstracts:
www.lcms2003.org/abstracts
- September 10 is the deadline for EARLY registration ; register now
before
prices go up: www.lcms2003.org/onlineReg.htm
Go to www.lcms2003.org/scientificProgram.htm
to view the updated
scientific program. The presenter’s list will be updated every week.
As a service to our members, we post Employment postings.
As a trial service to our members, we are now posting Situations Wanted. (Note: BAMS isn't a career center and will only post brief positions wanted for BAMS members only).
The San Francisco Bay Area Mass Spectrometry discussion group was formed in 1980 to provide a regular gathering for people interested in mass spectrometry and allied topics. BAMS currently has a membership of about 280 individual and 20 corporate members, and meets 8-10 times per year for a midweek dinner and lecture. Meetings attract between 30 and 90 people, and are held at a restaurant or hotel in the bay area convenient for our speaker. We usually convene at 6:00 pm for cocktails, dinner at 7:00 pm, and lecture at 8:15 pm.
If you have suggestions for a meeting topic, or know of a prominent mass spectroscopist who will be visiting the bay area, please contact our program chair.
To become a member, please fill out and mail in a membership
form.
BAMS has been fortunate to have had many excellent
speakers in our short history.
If you have suggestions for a meeting topic please contact our program
chair.
If you have questions or comments about BAMS, please contact one of
the BAMS officers.
Please thank our corporate sponsors who
help keep BAMS membership and dinner costs low.
Here are some internet links that may be of
interest to BAMS members.
If you have comments or suggestions for this web site, please email me at hugh-gregg@llnl.gov
Last update: 8/14/2003