Novel peptide recognized by RhoA GTPase

• Authors: Dominika Drulis-Fajdasz , Filip Jelen , Arkadiusz Oleksy , Jacek Otlewski
• Languages of publication: 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.

Keywords

EN

GrafGAP; RhoA GTPase; epitope mapping; PDZRhoGEF; peptide phage display

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2006
• Volume: 53
• Issue: 3
• Pages: 515-524

Dates

published

2006

received

2006-06-04

revised

2006-08-29

accepted

2006-09-04

(unknown)

2006-10-01

Contributors

author

Dominika Drulis-Fajdasz

• Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland

author

Filip Jelen

• Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland

author

Arkadiusz Oleksy

• Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland

author

Jacek Otlewski

• Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland

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