Degradation Parameters from Pulse-Chase Experiments

PLoS One. 2016 May 16;11(5):e0155028. doi: 10.1371/journal.pone.0155028. eCollection 2016.

Abstract

Pulse-chase experiments are often used to study the degradation of macromolecules such as proteins or mRNA. Considerations for the choice of pulse length include the toxicity of the pulse to the cell and maximization of labeling. In the general case of non-exponential decay, varying the length of the pulse results in decay patterns that look different. Analysis of these patterns without consideration to pulse length would yield incorrect degradation parameters. Here we propose a method that constructively includes pulse length in the analysis of decay patterns and extracts the parameters of the underlying degradation process. We also show how to extract decay parameters reliably from measurements taken during the pulse phase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Biosensing Techniques*
  • Computer Simulation
  • Kinetics
  • Markov Chains
  • Models, Biological
  • Proteins / metabolism*
  • Proteolysis*
  • RNA Stability*
  • RNA, Messenger / metabolism*

Substances

  • Proteins
  • RNA, Messenger

Grants and funding

CS was financially supported by the FP7 Marie-Curie Initial Training Networks program “Network for Integrated Cellular Homeostasis (NICHE)” grant number PITN-GA-2011-289384 (to AV). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.