High-precision mapping of protein protein interfaces: an integrated genetic strategy combining en masse mutagenesis and DNA-level parallel analysis on a yeast two-hybrid platform.
Pajunen, Maria ; Turakainen, Hilkka ; Poussu, Eini ; Peränen, Johan ; Vihinen, Mauno LU
and Savilahti, Harri
(2007)
In Nucleic Acids Research
35(16).
p.103-103
- Abstract
- Understanding networks of protein-protein interactions constitutes an essential component on a path towards comprehensive description of cell function. Whereas efficient techniques are readily available for the initial identification of interacting protein partners, practical strategies are lacking for the subsequent high-resolution mapping of regions involved in protein-protein interfaces. We present here a genetic strategy to accurately map interacting protein regions at amino acid precision. The system is based on parallel construction, sampling and analysis of a comprehensive insertion mutant library. The methodology integrates Mu in vitro transposition-based random pentapeptide mutagenesis of proteins, yeast two-hybrid screening and... (More)
- Understanding networks of protein-protein interactions constitutes an essential component on a path towards comprehensive description of cell function. Whereas efficient techniques are readily available for the initial identification of interacting protein partners, practical strategies are lacking for the subsequent high-resolution mapping of regions involved in protein-protein interfaces. We present here a genetic strategy to accurately map interacting protein regions at amino acid precision. The system is based on parallel construction, sampling and analysis of a comprehensive insertion mutant library. The methodology integrates Mu in vitro transposition-based random pentapeptide mutagenesis of proteins, yeast two-hybrid screening and high-resolution genetic footprinting. The strategy is general and applicable to any interacting protein pair. We demonstrate the feasibility of the methodology by mapping the region in human JFC1 that interacts with Rab8A, and we show that the association is mediated by the Slp homology domain 1. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/3635266
- author
- Pajunen, Maria
; Turakainen, Hilkka
; Poussu, Eini
; Peränen, Johan
; Vihinen, Mauno
LU
and Savilahti, Harri
- publishing date
- 2007
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Saccharomyces cerevisiae: genetics, Insertional: methods, Mutagenesis, Membrane Proteins: metabolism, Membrane Proteins: chemistry, Membrane Proteins: genetics, rab GTP-Binding Proteins: chemistry, rab GTP-Binding Proteins: metabolism
- in
- Nucleic Acids Research
- volume
- 35
- issue
- 16
- pages
- 103 - 103
- publisher
- Oxford University Press
- external identifiers
-
- pmid:17702760
- scopus:34548743990
- pmid:17702760
- ISSN
- 1362-4962
- DOI
- 10.1093/nar/gkm563
- language
- English
- LU publication?
- no
- id
- bef520fc-d4bb-48c8-8492-057ddf6f8097 (old id 3635266)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/17702760?dopt=Abstract
- date added to LUP
- 2016-04-04 08:09:35
- date last changed
- 2022-02-13 05:54:13
@article{bef520fc-d4bb-48c8-8492-057ddf6f8097,
abstract = {{Understanding networks of protein-protein interactions constitutes an essential component on a path towards comprehensive description of cell function. Whereas efficient techniques are readily available for the initial identification of interacting protein partners, practical strategies are lacking for the subsequent high-resolution mapping of regions involved in protein-protein interfaces. We present here a genetic strategy to accurately map interacting protein regions at amino acid precision. The system is based on parallel construction, sampling and analysis of a comprehensive insertion mutant library. The methodology integrates Mu in vitro transposition-based random pentapeptide mutagenesis of proteins, yeast two-hybrid screening and high-resolution genetic footprinting. The strategy is general and applicable to any interacting protein pair. We demonstrate the feasibility of the methodology by mapping the region in human JFC1 that interacts with Rab8A, and we show that the association is mediated by the Slp homology domain 1.}},
author = {{Pajunen, Maria and Turakainen, Hilkka and Poussu, Eini and Peränen, Johan and Vihinen, Mauno and Savilahti, Harri}},
issn = {{1362-4962}},
keywords = {{Saccharomyces cerevisiae: genetics; Insertional: methods; Mutagenesis; Membrane Proteins: metabolism; Membrane Proteins: chemistry; Membrane Proteins: genetics; rab GTP-Binding Proteins: chemistry; rab GTP-Binding Proteins: metabolism}},
language = {{eng}},
number = {{16}},
pages = {{103--103}},
publisher = {{Oxford University Press}},
series = {{Nucleic Acids Research}},
title = {{High-precision mapping of protein protein interfaces: an integrated genetic strategy combining en masse mutagenesis and DNA-level parallel analysis on a yeast two-hybrid platform.}},
url = {{http://dx.doi.org/10.1093/nar/gkm563}},
doi = {{10.1093/nar/gkm563}},
volume = {{35}},
year = {{2007}},
}