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NIDA Proteomics Center
> Keck Laboratory
Keck Foundation Biotechnology
Resource Laboratory at Yale University
The
Keck
Laboratory was founded in 1980 by Kenneth Williams, Ph.D. to bring a
wide range of state-of-the-art genomic and proteomic biotechnologies within
reach of hundreds of Yale and non-Yale investigators whose research programs
would otherwise not benefit from the highly sophisticated and expensive
instrumentation upon which biochemical and biomedical research is increasingly
dependent. To maximize its positive impact, the Keck Laboratory strives to
provide as many high quality services to as many investigators as possible.
Although priority is given to requests from Yale investigators, the decision to
also accept requests from scientists around the world ensures that when demand
by Yale investigators (which can be highly variable) for a particular
biotechnology is less than is needed to maintain maximal throughput, a backlog
of requests will be available to maintain high productivity. This policy, which
benefits all users of this Laboratory, minimizes operating costs and maximizes
the contribution of the Keck Laboratory and of Yale University to biochemical
and biomedical research. In keeping with this 25-year guiding philosophy, the
majority of analyses and syntheses are provided on a non-collaborative basis -
as it would not be feasible for this unit to collaborate simultaneously with
even a small fraction of the >1,100 investigators who utilize its services
annually. Throughout its history, the Keck Laboratory has been close to
self-supporting in that it has recovered >95% of its salary, supply, and
instrumentation costs from service charges, grants, contracts, and gifts.
Indeed, the Keck Laboratory space in the Boyer Center for Molecular Medicine was
constructed with a generous grant from the Keck Foundation - from whence this
unit derives its name. With 45 full time staff, including 12 with Ph.D. and 6
with M.S. level degrees, and >75 major instrument systems purchased at a cost of
$11 million dollars, the Keck Laboratory is one of the largest academic
biotechnology resource laboratories of its kind in the world.
The 12 individual Resources that are the Keck
Laboratory provide a wide range of genomic and proteomic syntheses and analyses
that include DNA microarray and Affymetrix gene chip technology, oligo and
peptide synthesis, DNA and protein sequencing, biophysical analysis of proteins
and other biopolymers (including size exclusion HPLC/laser light scattering and
stopped flow fluorescence/absorbance), biostatistical analyses, and mass
spectrometry (MS). DNA microarray technology is one of the newer genomic
technologies offered and the Keck Lab has available 1" x 3" glass slide
microarrays that allow the relative level of expression of 27,000 Arabidopsis,
16,000 mouse, 5,000 rat or 21,000 human genes and ESTs to be interrogated in
parallel in a single experiment. The rapid growth of the DNA Microarray Resource
has been aided considerably by support received from its NIDDK Microarray
Biotechnology Center Grant, which is one of 16 centers nationally.
Although protein profiling technologies lag far
behind that of mRNA expression analysis, the Keck Laboratory has initiated three
protein profiling technologies, MALDI-MS based peptide/protein disease biomarker
discovery, isotope-coded affinity tag (ICAT)/MS based protein profiling, and
differential fluorescence 2D gel electrophoresis (DIGE). The Amersham system
that is the basis for the DIGE technology includes a gel picker that can
automatically excise and transfer to 96 well plates spots whose relative level
of differential expression in a control versus experimental sample meets
user-defined criteria (e.g., >3-fold change). Typically, more than 50 of the
1,000 or more protein-containing spots obtained from the analysis of a complex
tissue protein extract are selected for protein identification from each DIGE
gel. Instrumentation also has been installed recently to support automated 2D
chromatofocusing/RP-HPLC based-protein profiling and the Keck Laboratory was
awarded a $1.5 million NIH High End Instrumentation Grant to purchase a Bruker
9.4T FTICR mass spectrometer that will be installed in June, 2004. Major uses
envisioned for this state-of-the-art MS platform are multiple dimension protein
identification technology (MudPIT) and phospho-proteome profiling. Indicative of
the increasing need for higher throughput technologies, the Keck Laboratory
initiated an automated, LC-MS/MS-based protein identification service that
starts with investigators submitting protein-containing, polyacrylamide gel
bands or spots (i.e., from DIGE analyses). The Keck Laboratory also provides a
single nucleotide polymorphism (SNP) genotyping service with a maximum
throughput of >20,000 SNP genotypes/day and that is based on MS analysis of the
products of single base extension reactions.
As suggested by the number of services it
provides, the Keck Laboratory is making an important contribution to biological
and biomedical research that extends far beyond Yale University. In 2002 the
Keck Lab provided 215,383 genomic and proteomic analyses and syntheses to 422
Yale and 692 non-Yale investigators at more than 350 institutions in 20
countries. To enable the Keck Lab to continue to expand to meet the 40% average
annual increase (since 1990) in demand for services, the Yale School of Medicine
has provided additional space that has brought the total Keck Laboratory space
to approximately 25,000 ft2 by July, 2004. This space includes a new, 5,425 ft2
custom-designed mass spectrometry laboratory. To help its users take maximum
advantage of its Resources, the
Keck web
pages provide extensive background information on the biotechnologies it
offers and on interpreting the resulting data. That this effort is succeeding is
suggested by the web log which indicates, for instance, that in July, 2004 the
Keck web pages received 870,923 requests from 6,726 distinct hosts.
The Keck Laboratory contains and/or is very
closely associated with three major centers:
Yale/NHLBI Proteomics Center
Northeast Biodefense Center (NBC)
One of 8 Regional Centers of Excellence
established in 2003.
NBC consists of 200 faculty at the CDC and 36
institutions in NY, CT, and NJ.
NBC Proteomics Core is within the Keck
Laboratory’s MS Resource. It supports:
Yale/NIDA Neuroproteomics Research Center
The theme for this Center is "Proteomics of
Altered Signaling in Addiction".
The Center brings together exceptionally strong
Yale programs in proteomics and signal transduction in the brain to identify
adaptive changes in protein signaling that occur in response to substance abuse.
Major areas of signal transduction research that
are being studied in the Center and that we believe represent closely
intertwined research areas are:
Receptor-mediated signaling, with a primary focus
on protein phosphorylation, but also an increasing interest in signaling
mediated by lipid phosphorylation and metabolism
Signaling in the regulation of membrane traffic:
synaptic vesicle exo-endocytosis presynaptically and of the traffic of
neurotransmitter receptors postsynaptically
Signaling and the regulation of synaptic structure
Signaling and the regulation of transcription
As judged by extensive use of the Keck Laboratory
by non-Yale investigators and by comments like the following by the NIH Study
Panel that reviewed its most recent Shared Instrumentation Grant (RR151837 in
2001), the Keck Laboratory provides a very high level of service to its users
and is dedicated to continuing to do so far into the future:
"This is an outstanding proposal from a
premiere core laboratory at Yale."
"The
Keck Biotechnology Resource Laboratory is arguably one of the best facilities of
its kind in the nation in terms of support and expertise. The large number of
researchers who annually make use of this facility, both from the Yale campus
and nationwide, is testament to the significance of the facility for crucial
support of biomedical and biotechnology research."
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