Biochemical characterization of arabidopsis developmentally regulated G-proteins (DRGs)

O'Connell, A, Robin, G, Kobe, B and Botella, JR (2009) Biochemical characterization of arabidopsis developmentally regulated G-proteins (DRGs). PROTEIN EXPRESSION AND PURIFICATION, 67 2: 88-95. doi:10.1016/j.pep.2009.05.009


Author O'Connell, A
Robin, G
Kobe, B
Botella, JR
Title Biochemical characterization of arabidopsis developmentally regulated G-proteins (DRGs)
Journal name PROTEIN EXPRESSION AND PURIFICATION   Check publisher's open access policy
ISSN 1046-5928
Publication date 2009-10-01
Year available 2009
Sub-type Article (original research)
DOI 10.1016/j.pep.2009.05.009
Volume 67
Issue 2
Start page 88
End page 95
Total pages 8
Editor Burgess, R.R.
Place of publication United States
Publisher Academic Press
Collection year 2010
Language eng
Subject C1
060702 Plant Cell and Molecular Biology
8298 Environmentally Sustainable Plant Production
Abstract Developmentally regulated G-proteins (DRGs) are a highly conserved family of GTP-binding proteins found in archaea, plants, fungi and animals, indicating important roles in fundamental pathways. Their function is poorly understood, but they have been implicated in cell division, proliferation, and growth, as well as several medical conditions. Individual subfamilies within the G-protein superfamily possess unique nucleotide binding and hydrolysis rates that are intrinsic to their cellular function, and so characterization of these rates for a particular G-protein may provide insight into its cellular activity. We have produced recombinant active DRG protein using a bacterial expression system and refolding, and performed biochemical characterization of their GTP binding and hydrolysis. We show that recombinant Arabidopsis thaliana atDRG1 and atDRG2a are able to bind GDP and GTP. We also show that DRGs can hydrolyze GTP in vitro without the assistance of GTPase-activating proteins and guanine exchange factors. The atDRG proteins hydrolyze GTP at a relatively slow rate (0.94 103 min1 for DRG1 and 1.36 103 min1 for DRG2) that is consistent with their nearest characterized relatives, the Obg subfamily. The ability of DRGs to bind nucleotide substrates without assistance, their slow rate of GTP hydrolysis, heat stress activation and domain conservation suggest a possible role as a chaperone in ribosome assembly in response to stress as it has been suggested for the Obg proteins, a different but related G-protein subfamily.
Keyword Developmentally regulated G-protein
GTP-binding protein
DRG
Arabidopsis thaliana
GTP-BINDING PROTEIN
BACILLUS-SUBTILIS
SECONDARY STRUCTURE
OBG
SPORULATION
EXPRESSION
REVEALS
GENEVESTIGATOR
INITIATION
SIGMA(B)
Q-Index Code C1
Q-Index Status Confirmed Code

 
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Created: Sun, 24 Jan 2010, 10:00:43 EST