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Related Concept Videos

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An experimental design is a systematic process that allows researchers to evaluate the relationship between dependent and independent variables. There are three widely used types of experimental design - pre-experimental design, true experimental design, and quasi-experimental design. In pre-experimental design, the researcher compares the data before and after some interventions or treatments. The true-experimental design has more than one purposefully created group, a commonly measured...
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The time response of a linear time-invariant (LTI) system can be divided into transient and steady-state responses. The transient response represents the system's initial reaction to a change in input and diminishes to zero over time. In contrast, the steady-state response is the behavior that persists after the transient effects have faded.
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Characterization of Complex Systems Using the Design of Experiments Approach: Transient Protein Expression in Tobacco as a Case Study
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Community-Wide Experimental Evaluation of the PROSS Stability-Design Method.

Yoav Peleg1, Renaud Vincentelli2, Brett M Collins3

  • 1Department of Life Sciences Core Facilities (LSCF), Weizmann Institute of Science, Rehovot 7610001, Israel.

Journal of Molecular Biology
|March 30, 2021
PubMed
Summary
This summary is machine-generated.

The PROSS algorithm enhances protein stability and expression, even for challenging targets. This study shows its effectiveness as a user-friendly tool for protein scientists, requiring tailored conditions for optimal results.

Keywords:
PROSSProtein expressionProtein stabilityRecombinant proteinsRosetta

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Area of Science:

  • Protein Engineering
  • Computational Biology
  • Biotechnology

Background:

  • Protein design methods have advanced, but often require expert knowledge.
  • The PROSS algorithm aims to improve protein stability and expression levels.
  • Widespread adoption of protein design tools is limited by complexity.

Purpose of the Study:

  • To benchmark the PROSS algorithm as a stand-alone tool for protein scientists with limited modeling experience.
  • To assess the general applicability of PROSS across diverse protein targets.
  • To evaluate PROSS designs for expression, stability, and activity.

Main Methods:

  • PROSS algorithm applied to 14 unrelated protein targets by 12 laboratories.
  • Designs evaluated for heterologous expression in prokaryotic and eukaryotic systems.
  • Thermal stability and activity assays performed on selected targets.

Main Results:

  • Increased heterologous expression observed in 9 out of 14 targets.
  • Improved thermal stability noted in 9 out of 10 tested targets.
  • PROSS designs enabled expression of previously unexpressible proteins (hSCF, CLD12).

Conclusions:

  • PROSS can enhance stability and expressibility in diverse proteins.
  • Target-specific expression conditions are typically required for optimal PROSS performance.
  • Community-wide benchmarking demonstrates PROSS's utility and identifies areas for future algorithm development.