Scientific Report 2007
Mass Spectrometry for Analysis of Proteins
J.A. Caldwell Busby, V. Cavett
work to develop methods compatible with mass spectrometry for the analysis of proteins.
Our areas of interest are wide-ranging and include methods for subcellular compartmentalization
for large-scale proteomics profiling and identification of proteins involved in
gene regulation. Gene regulation, or epigenomics, is of particular interest. To
date, the focus of epigenomics research has been patterns of DNA methylation and
posttranslational modifications of the histone N termini. However, many other proteins
are associated with chromatin, including transcription factors, enzymes, and scaffolding
proteins. We are developing methods for isolating and analyzing the supermolecular
structure of chromatin, particularly the regulatory protein machineries, and we
are using cutting-edge mass spectrometric techniques to characterize the proteins
involved in gene regulation.
Study of the
interactions of proteins and DNA biopolymers is not new; much work has been done
on transcription and gene regulation. However, most of this research has relied
on the analysis of DNA methylation patterns and chromatin immunoprecipitation methods.
Although informative, these studies ignore the extensive protein components of the
system, potentially a rich source of information about how epigenetic patterns are
established, maintained, and modified in healthy tissue. Such studies also cannot
be used to assess how changes in these protein components contribute to disease
states, aging, or cell death.
is a widely used technique in proteomics. Powerful, high-resolution instruments,
coupled with new fragmentation methods, including electron capture dissociation,
are driving the identification of novel proteins and new sites of protein identification.
This identification is something for which neither traditional molecular biology
nor modern RNA/DNA analysis is well suited. The development of a mass spectrometry–based
process to identify components of the chromatin regulatory network will yield a
broadly applicable method that will contribute to a wide range of biological investigations,
including chromatin regulation of gene expression.
M.D., Sayyah, J., VonDerLinden, D., Johns, M., Ostrov, D.A., Caldwell-Busby, J.,
Sayeski, P.P. The N-terminal
SH2 domain of the tyrosine phosphatase, SHP-2, is essential for Jak2-dependent signaling
via angiotensin II type AT1 receptor. Cell Signal. 19:600, 2007.
S., Smith, S., Algeciras, M., Cavett, V., Caldwell Busby, J.A., London, S., Clayton,
D.F., Bhattacharya, S.K.
Proteomic analyses of songbird (zebra finch; Taeniopygia guttata) retina.
J. Proteome Res. 6:1093, 2007.
S., Smith, S., Gandhi, S., Caldwell Busby, J.A., London, S., Luksch, H., Clayton,
D.F., Bhattacharya, S.K. Proteomic
analyses of zebra finch optic tectum and comparative histochemistry. J. Proteome
Res. 6:2341, 2007.