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Yeast Signaling01:28

Yeast Signaling

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Yeasts are single-celled organisms, but unlike bacteria, they are eukaryotes (cells with a nucleus). Cell signaling in yeast is similar to signaling in other eukaryotic cells. A ligand, such as a protein or a small molecule released from a yeast cell, attaches to a receptor on the cell surface. The binding stimulates second-messenger kinases to activate or inactivate transcription factors that further regulate gene expression. Many of the yeast intracellular signaling cascades have similar...
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Protein Engineering by Yeast Surface Display
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Yeast Surface Display for Protein Engineering: Library Generation, Screening, and Affinity Maturation.

Byong H Kang1,2, Brianna M Lax3,2, K Dane Wittrup4,5,6

  • 1Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|April 28, 2022
PubMed
Summary
This summary is machine-generated.

Yeast surface display enables protein engineering for desired traits. Updated protocols detail screening large libraries for binders and improving their affinity through iterative mutagenesis and cell sorting.

Keywords:
Affinity maturationDirected evolutionLibrary screeningProtein engineeringYeast surface display

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Area of Science:

  • Biotechnology and Molecular Biology
  • Protein Engineering
  • Directed Evolution

Background:

  • Yeast surface display is a versatile platform for protein engineering.
  • Directed evolution strategies are crucial for optimizing protein properties.
  • Identifying and refining protein binders is essential in biotechnology.

Purpose of the Study:

  • To present updated protocols for yeast surface display.
  • To detail methods for identifying lead binders.
  • To describe affinity maturation techniques for engineered proteins.

Main Methods:

  • Screening large libraries using magnetic bead selections.
  • Employing flow cytometric selections for binder identification.
  • Iterative cycles of mutagenesis and fluorescence-activated cell sorting for affinity improvement.

Main Results:

  • Successful identification of lead protein binders from large libraries.
  • Demonstrated improvement in protein affinities through the described methods.
  • Characterization of single clones for desired properties.

Conclusions:

  • Yeast surface display provides an effective framework for protein engineering.
  • The presented protocols enhance the identification and optimization of protein binders.
  • This methodology facilitates the development of proteins with improved functionalities.