1. Academic Validation
  2. Chimeric proteins tagged with specific 3xHA cassettes may present instability and functional problems

Chimeric proteins tagged with specific 3xHA cassettes may present instability and functional problems

  • PLoS One. 2017 Aug 11;12(8):e0183067. doi: 10.1371/journal.pone.0183067.
Sara Saiz-Baggetto 1 Ester Méndez 1 Inma Quilis 1 J Carlos Igual 1 M Carmen Bañó 1
Affiliations

Affiliation

  • 1 Departament de Bioquímica i Biologia Molecular and Estructura de Recerca Interdisciplinar en Biotecnologia i Biomedicina, Universitat de València, Burjassot (Valencia), Spain.
Abstract

Epitope-tagging of proteins has become a widespread technique for the analysis of protein function, protein interactions and protein localization among Others. Tagging of genes by chromosomal integration of PCR amplified cassettes is a widely used and fast method to label proteins in vivo. Different systems have been developed during years in the yeast Saccharomyces cerevisiae. In the present study, we analysed systematically a set of yeast proteins that were fused to different tags. Analysis of the tagged proteins revealed an unexpected general effect on protein level when some specific tagging module was used. This was due in all cases to a destabilization of the proteins and caused a reduced protein activity in the cell that was only apparent in particular conditions. Therefore, an extremely cautious approach is required when using this strategy.

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Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-P5491
    99.08%, Bioactive Peptide