Biochemical clustering of monomeric GTPases of the Ras superfamily

被引:14
作者
Caruso, ME
Jenna, S
Beaulne, S
Lee, EH
Bergeron, A
Chauve, C
Roby, P
Rual, JF
Hill, DE
Vidal, M
Bossé, R
Chevet, E [1 ]
机构
[1] PerkinElmer BioSignal, Montreal, PQ H3J 1R4, Canada
[2] McGill Univ, Dept Surg, Organelle Signaling Lab, Montreal, PQ H3A 2B2, Canada
[3] McGill Univ, Montreal Proteom Network, Montreal, PQ H3A 2B2, Canada
[4] Univ Quebec, Lab Combinatoire & Informat Math, Montreal, PQ H3C 3P8, Canada
[5] Dana Farber Canc Inst, Ctr Canc Syst Biol, Boston, MA 02115 USA
[6] Dana Farber Canc Inst, Dept Canc Biol, Boston, MA 02115 USA
[7] Harvard Univ, Sch Med, Dept Genet, Boston, MA 02115 USA
关键词
D O I
10.1074/mcp.M500025-MCP200
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
To date phylogeny has been used to compare entire families of proteins based on their nucleotide or amino acid sequence. Here we developed a novel analytical platform allowing a systematic comparison of protein families based on their biochemical properties. This approach was validated on the Rho subfamily of GTPases. We used two high throughput methods, referred to as AlphaScreen(TM) and FlashPlate(R), to measure nucleotide binding capacity, exchange, and hydrolysis activities of small monomeric GTPases. These two technologies have the characteristics to be very sensitive and to allow homogenous and high throughput assays. To analyze and integrate the data obtained, we developed an algorithm that allows the classification of GTPases according to their enzymatic activities. Integration and hierarchical clustering of these results revealed unexpected features of the small Rho GTPases when compared with primary sequence-based trees. Hence we propose a novel phylobiochemical classification of the Ras superfamily of GTPases.
引用
收藏
页码:936 / 944
页数:9
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