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2006 | 53 | 3 | 515-524
Article title

Novel peptide recognized by RhoA GTPase

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EN
Abstracts
EN
A phage-displayed random 7-mer disulfide bridge-constrained peptide library was used to map the surface of the RhoA GTPase and to find peptides able to recognize RhoA switch regions. Several peptide sequences were selected after four rounds of enrichment, giving a high signal in ELISA against RhoA-GDP. A detailed analysis of one such selected peptide, called R2 (CWSFPGYAC), is reported. The RhoA - R2 interaction was investigated using fluorescence spectroscopy, chemical denaturation, and determination of the kinetics of nucleotide exchange and GTP hydrolysis in the presence of RhoA regulatory proteins. All measurements indicate that the affinity of the R2 peptide for RhoA is in the micromolar range and that R2 behaves as an inhibitor of: i) GDP binding to the apo form of RhoA (Mg2+- and nucleotide-free form of the GTPase), ii) nucleotide exchange stimulated by GEF (DH/PH tandem from PDZRhoGEF), and iii) GTP hydrolysis stimulated by the BH domain of GrafGAP protein.
Publisher

Year
Volume
53
Issue
3
Pages
515-524
Physical description
Dates
published
2006
received
2006-06-04
revised
2006-08-29
accepted
2006-09-04
(unknown)
2006-10-01
Contributors
  • Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland
author
  • Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland
  • Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland
  • Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland
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Document Type
Publication order reference
Identifiers
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv53p515kz
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