In Vivo Reporters for Protein Half-Life

Methods Mol Biol. 2017:1669:387-406. doi: 10.1007/978-1-4939-7286-9_29.

Abstract

Determination of the general capacity of proteolytic activity of a certain cell or tissue type can be crucial for an assessment of various features of an organism's growth and development and also for the optimization of biotechnological applications. Here, we describe the use of chimeric protein stability reporters that can be detected by standard laboratory techniques such as histological staining, selection using selective media or fluorescence microscopy. Dependent on the expression of the reporters due to the promoters applied, cell- and tissue-specific questions can be addressed. Here, we concentrate on methods which can be used for large-scale screening for protein stability changes rather than for detailed protein stability studies.

Keywords: Histology; N-end rule; Protein degradation; Proteostasis; Ubiquitination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Microscopy, Fluorescence
  • Plant Proteins / chemistry
  • Plant Proteins / metabolism*
  • Proteolysis
  • Ubiquitination

Substances

  • Plant Proteins