Biochemical clustering of monomeric GTPases of the Ras superfamily.
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 and FlashPlate, 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.
Caruso
Marie-Elaine
ME
Organelle Signaling Laboratory, Department of Surgery and Montreal Proteomics Network, McGill University, Montreal, Quebec H3A 2B2, Canada.
Jenna
Sarah
S
Beaulne
Simon
S
Lee
Eun-Hye
EH
Bergeron
Anne
A
Chauve
Cédric
C
Roby
Philippe
P
Rual
Jean-François
JF
Hill
David E
DE
Vidal
Marc
M
Bossé
Roger
R
Chevet
Eric
E
D000465:Algorithms; D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000201:enzymology / Q000235:genetics; D016000:Cluster Analysis; D049751:Genome, Helminth; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010802:Phylogeny; D011485:Protein Binding; D018631:ras Proteins / Q000145:classification / Q000378:metabolism*; D020741:rho GTP-Binding Proteins / Q000145:classification / Q000378:metabolism*
D018631:ras Proteins; D020741:rho GTP-Binding Proteins