Yeast display
Encyclopedia
Yeast display is a technique used in the field of protein engineering
. The yeast display technique was first published by the laboratory
of Professor K. Dane Wittrup. The technology was sold to Abbott Laboratories in 2001.
A protein of interest is displayed as a fusion to the Aga2p protein on the surface of yeast
. The Aga2p protein is naturally used by yeast to mediate cell–cell contacts during yeast cell mating. As such, display of a protein via Aga2p projects the protein away from the cell surface, minimizing potential interactions with other molecules on the yeast cell wall. The use of magnetic separation and flow cytometry
in conjunction with a yeast display library is a highly effective method to isolate high affinity protein
ligand
s against nearly any receptor
through directed evolution
.
Disadvantages include smaller mutant library sizes compared to alternative methods and differential glycosylation
in yeast compared to mammalian cells. These disadvantages have not limited the success of yeast display for a number of applications, including engineering the highest monovalent ligand-binding affinity reported to date for an engineered protein (Boder, E.T. 2000)
Alternative methods for protein evolution in vitro are phage display
, ribosome display
, bacterial display
, and mRNA display
.
Protein engineering
Protein engineering is the process of developing useful or valuable proteins. It is a young discipline, with much research taking place into the understanding of protein folding and recognition for protein design principles....
. The yeast display technique was first published by the laboratory
Laboratory
A laboratory is a facility that provides controlled conditions in which scientific research, experiments, and measurement may be performed. The title of laboratory is also used for certain other facilities where the processes or equipment used are similar to those in scientific laboratories...
of Professor K. Dane Wittrup. The technology was sold to Abbott Laboratories in 2001.
A protein of interest is displayed as a fusion to the Aga2p protein on the surface of yeast
Yeast
Yeasts are eukaryotic micro-organisms classified in the kingdom Fungi, with 1,500 species currently described estimated to be only 1% of all fungal species. Most reproduce asexually by mitosis, and many do so by an asymmetric division process called budding...
. The Aga2p protein is naturally used by yeast to mediate cell–cell contacts during yeast cell mating. As such, display of a protein via Aga2p projects the protein away from the cell surface, minimizing potential interactions with other molecules on the yeast cell wall. The use of magnetic separation and flow cytometry
Flow cytometry
Flow cytometry is a technique for counting and examining microscopic particles, such as cells and chromosomes, by suspending them in a stream of fluid and passing them by an electronic detection apparatus. It allows simultaneous multiparametric analysis of the physical and/or chemical...
in conjunction with a yeast display library is a highly effective method to isolate high affinity protein
Protein
Proteins are biochemical compounds consisting of one or more polypeptides typically folded into a globular or fibrous form, facilitating a biological function. A polypeptide is a single linear polymer chain of amino acids bonded together by peptide bonds between the carboxyl and amino groups of...
ligand
Ligand (biochemistry)
In biochemistry and pharmacology, a ligand is a substance that forms a complex with a biomolecule to serve a biological purpose. In a narrower sense, it is a signal triggering molecule, binding to a site on a target protein.The binding occurs by intermolecular forces, such as ionic bonds, hydrogen...
s against nearly any receptor
Receptor (biochemistry)
In biochemistry, a receptor is a molecule found on the surface of a cell, which receives specific chemical signals from neighbouring cells or the wider environment within an organism...
through directed evolution
Directed evolution
thumb|250px|right|An example of a possible round to evolve a protein based fluorescent sensor for a specific analyte using two consecutive FACS sortings...
.
Advantages and disadvantages of yeast display
Advantages of yeast display over other in vitro evolution methods include eukaryotic expression and processing, quality control mechanisms of the eukaryotic secretory pathway, minimal avidity effects, and quantitative library screening through fluorescent-activated cell sorting (FACS).Disadvantages include smaller mutant library sizes compared to alternative methods and differential glycosylation
Glycosylation
Glycosylation is the reaction in which a carbohydrate, i.e. a glycosyl donor, is attached to a hydroxyl or other functional group of another molecule . In biology glycosylation refers to the enzymatic process that attaches glycans to proteins, lipids, or other organic molecules...
in yeast compared to mammalian cells. These disadvantages have not limited the success of yeast display for a number of applications, including engineering the highest monovalent ligand-binding affinity reported to date for an engineered protein (Boder, E.T. 2000)
Alternative methods for protein evolution in vitro are phage display
Phage display
Phage display is a method for the study of protein–protein, protein–peptide, and protein–DNA interactions that uses bacteriophages to connect proteins with the genetic information that encodes them. Phage Display was originally invented by George P...
, ribosome display
Ribosome display
Ribosome display is a technique used to perform in vitro protein evolution to create proteins that can bind to a desired ligand. The process results in translated proteins that are associated with their mRNA progenitor which is used, as a complex, to bind to an immobilized ligand in a selection step...
, bacterial display
Bacterial display
Bacterial display is a protein engineering technique used for in vitro protein evolution...
, and mRNA display
MRNA display
mRNA display is a display technique used for in vitro protein, and/or peptide evolution to create molecules that can bind to a desired target. The process results in translated peptides or proteins that are associated with their mRNA progenitor via a puromycin linkage. The complex then binds to...
.