Application of Mu in vitro transposition for high-precision mapping of protein-protein interfaces on a yeast two-hybrid platform

Maria Pajunen, Eini Poussu, Hilkka Turakainen, Harri Savilahti

    Research output: Contribution to journalArticleScientificpeer-review

    Abstract

    High-precision mapping of regions involved in protein-protein interfaces of interacting protein partners is an essential component on a path to understand various cellular functions. Trans poson-based systems, particularly those involving in vitro reactions, offer exhaustive insertion mutant libraries and high-throughput platforms for many types of genetic analyses. We present here a genetic strategy to accurately map interacting protein regions at amino acid precision that is based on transposition-assisted construction, sampling, and analysis of a comprehensive insertion mutant library. The methodology integrates random pentapeptide mutagenesis of proteins, yeast two-hybrid screening, and high-resolution genetic footprinting. This straightforward strategy is general, and it provides a rapid and easy means to identify critical contact regions in proteins without the requirement of prior structural knowledge. (C) 2009 Elsevier Inc. All rights reserved.
    Original languageEnglish
    JournalMethods
    Volume49
    Issue number3
    Pages (from-to)255-262
    Number of pages8
    ISSN1046-2023
    DOIs
    Publication statusPublished - 2009
    MoE publication typeA1 Journal article-refereed

    Cite this

    Pajunen, Maria ; Poussu, Eini ; Turakainen, Hilkka ; Savilahti, Harri. / Application of Mu in vitro transposition for high-precision mapping of protein-protein interfaces on a yeast two-hybrid platform. In: Methods. 2009 ; Vol. 49, No. 3. pp. 255-262.
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    abstract = "High-precision mapping of regions involved in protein-protein interfaces of interacting protein partners is an essential component on a path to understand various cellular functions. Trans poson-based systems, particularly those involving in vitro reactions, offer exhaustive insertion mutant libraries and high-throughput platforms for many types of genetic analyses. We present here a genetic strategy to accurately map interacting protein regions at amino acid precision that is based on transposition-assisted construction, sampling, and analysis of a comprehensive insertion mutant library. The methodology integrates random pentapeptide mutagenesis of proteins, yeast two-hybrid screening, and high-resolution genetic footprinting. This straightforward strategy is general, and it provides a rapid and easy means to identify critical contact regions in proteins without the requirement of prior structural knowledge. (C) 2009 Elsevier Inc. All rights reserved.",
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    Application of Mu in vitro transposition for high-precision mapping of protein-protein interfaces on a yeast two-hybrid platform. / Pajunen, Maria; Poussu, Eini; Turakainen, Hilkka; Savilahti, Harri.

    In: Methods, Vol. 49, No. 3, 2009, p. 255-262.

    Research output: Contribution to journalArticleScientificpeer-review

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    AU - Pajunen, Maria

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    AU - Turakainen, Hilkka

    AU - Savilahti, Harri

    PY - 2009

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    AB - High-precision mapping of regions involved in protein-protein interfaces of interacting protein partners is an essential component on a path to understand various cellular functions. Trans poson-based systems, particularly those involving in vitro reactions, offer exhaustive insertion mutant libraries and high-throughput platforms for many types of genetic analyses. We present here a genetic strategy to accurately map interacting protein regions at amino acid precision that is based on transposition-assisted construction, sampling, and analysis of a comprehensive insertion mutant library. The methodology integrates random pentapeptide mutagenesis of proteins, yeast two-hybrid screening, and high-resolution genetic footprinting. This straightforward strategy is general, and it provides a rapid and easy means to identify critical contact regions in proteins without the requirement of prior structural knowledge. (C) 2009 Elsevier Inc. All rights reserved.

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