Nature Genetics
26, 283 - 289 (2000)
doi:10.1038/81576
Analysis of yeast protein kinases using protein chipsHeng Zhu1, James F. Klemic2, 3, Swan Chang2, Paul Bertone1, Antonio Casamayor1, Kathryn G. Klemic4, David Smith1, Mark Gerstein5, Mark A. Reed2, 3
& Michael Snyder1, 51
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, Connecticut, USA. 2
Department of Electrical Engineering, Yale University, New Haven, Connecticut, USA. 3
Department of Applied Physics, Yale University, New Haven, Connecticut, USA. 4
Department of Cellular and Molecular Physiology, Yale University, New Haven, Connecticut, USA.
5
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, USA
.
Correspondence should be addressed to Michael Snyder michael.snyder@yale.eduWe have developed a novel protein chip technology that allows the high-throughput
analysis of biochemical activities, and used this approach to analyse nearly
all of the protein kinases from Saccharomyces cerevisiae. Protein chips
are disposable arrays of microwells in silicone elastomer sheets placed on
top of microscope slides. The high density and small size of the wells allows
for high-throughput batch processing and simultaneous analysis of many individual
samples. Only small amounts of protein are required. Of 122 known and predicted
yeast protein kinases, 119 were overexpressed and analysed using 17 different
substrates and protein chips. We found many novel activities and that a large
number of protein kinases are capable of phosphorylating tyrosine. The tyrosine
phosphorylating enzymes often share common amino acid residues that lie near
the catalytic region. Thus, our study identified a number of novel features
of protein kinases and demonstrates that protein chip technology is useful
for high-throughput screening of protein biochemical activity.
|
