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0  structures 18  species 0  interactions 18  sequences 1  architecture

Family: AGA2 (PF17366)

Summary: A-agglutinin-binding subunit Aga2

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This is the Wikipedia entry entitled "Yeast display". More...

Yeast display Edit Wikipedia article

Yeast display (or yeast surface display) is a protein engineering technique that uses the expression of recombinant proteins incorporated into the cell wall of yeast for isolating and engineering antibodies.[1]

Development

The yeast display technique was first published by the laboratory of Professor K. Dane Wittrup.[2] The technology was sold to Abbott Laboratories in 2001.[3]

How it works

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 ligands against nearly any receptor through directed evolution.

Advantages and disadvantages

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). Yeast are eukaryotic organisms that allow for complex post-translational modifications to proteins that no other display libraries are able to provide.

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) Currently the yeast display library created by Boder is no longer available, as the INVSc1 cell line from Invitrogen is no longer available due to IP issues.

Alternative methods for protein evolution in vitro are mammalian display, phage display, ribosome display, bacterial display, and mRNA display.

References

  1. ^ Gai, S Annie; Wittrup, K Dane (2007). "Yeast surface display for protein engineering and characterization". Current Opinion in Structural Biology. 17 (4): 467–473. doi:10.1016/j.sbi.2007.08.012. ISSN 0959-440X. 
  2. ^ Boder, Eric T.; Wittrup, K. Dane (1997). "Yeast surface display for screening combinatorial polypeptide libraries". Nature Biotechnology. 15 (6): 553–557. doi:10.1038/nbt0697-553. ISSN 1087-0156. 
  3. ^ http://www.news.uiuc.edu/NEWS/01/1221biodisplaytechnology.html

Further reading

  • Boder, E.T., Wittrup, K.D.; Biotechnol. Prog., 1998, 14, 55–62.
  • Boder E.T., Midelfort K.S., Wittrup K.D.; Proc Natl Acad Sci, 2000, 97(20):10701-10705.
  • Graff, C.P., Chester, K., Begent, R., Wittrup, K.D.; Prot. Eng. Des. Sel., 2004, 17, 293–304.
  • Feldhaus M, Siegel R.; Methods in Molecular Biology 263:311–332 (2004).
  • Weaver-Feldhaus, Jane M; Lou, Jianlong; Coleman, James R; Siegel, Robert W; Marks, James D; Feldhaus, Michael J (2004). "Yeast mating for combinatorial Fab library generation and surface display". FEBS Letters. 564 (1-2): 24–34. doi:10.1016/S0014-5793(04)00309-6. ISSN 0014-5793. 

This page is based on a Wikipedia article. The text is available under the Creative Commons Attribution/Share-Alike License.

This tab holds the annotation information that is stored in the Pfam database. As we move to using Wikipedia as our main source of annotation, the contents of this tab will be gradually replaced by the Wikipedia tab.

A-agglutinin-binding subunit Aga2 Provide feedback

The wall of Saccharomyces cerevisiae consists of mannoproteins, β-glucans, and a small amount of chitin [1]. Mannoproteins include Aga2p where this domain family is found. There are two main display systems for yeast, the agglutinin system and the flocculin system [2]. The S. cerevisiae sexual agglutinins facilitate the mating between two types of cells, a and α. a-Agglutinin consists of two subunits, encoded by two unlinked genes, AGA1 and AGA2 [3]. The cell surface adhesion protein (Aga2), enhances agglutination between a and α cells. Optimal binding includes interactions of the α-agglutinin binding pocket with the Aga2p terminal carboxyl group. This O-mannosylated glycopeptide is doubly disulfide linked to Aga1p. The Aga2p half-cystines near the ends of the peptide are linked to two Aga1p Cys residues separated by only two residues. This closeness of the disulfide bonds stabilizes the α/β structure in Aga2p [4].

Literature references

  1. Ananphongmanee V, Srisala J, Sritunyalucksana K, Boonchird C;, PLoS One. 2015;10:e0128764.: Yeast Surface Display of Two Proteins Previously Shown to Be Protective Against White Spot Syndrome Virus (WSSV) in Shrimp. PUBMED:26083446 EPMC:26083446

  2. Jahns AC, Rehm BH;, Microb Biotechnol. 2012;5:188-202.: Relevant uses of surface proteins--display on self-organized biological structures. PUBMED:21906264 EPMC:21906264

  3. Xie X, Lipke PN;, Yeast. 2010;27:479-488.: On the evolution of fungal and yeast cell walls. PUBMED:20641026 EPMC:20641026

  4. Dranginis AM, Rauceo JM, Coronado JE, Lipke PN; , Microbiol Mol Biol Rev. 2007;71:282-294.: A biochemical guide to yeast adhesins: glycoproteins for social and antisocial occasions. PUBMED:17554046 EPMC:17554046


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(16)
Full
(18)
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(36)
NCBI
(46)
Meta
(0)
RP15
(1)
RP35
(8)
RP55
(13)
RP75
(18)
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  Seed
(16)
Full
(18)
Representative proteomes UniProt
(36)
NCBI
(46)
Meta
(0)
RP15
(1)
RP35
(8)
RP55
(13)
RP75
(18)
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This family is new in this Pfam release.

Seed source: PRODOM:PD062190
Previous IDs: none
Type: Family
Author: El-Gebali S
Number in seed: 16
Number in full: 18
Average length of the domain: 58.80 aa
Average identity of full alignment: 41 %
Average coverage of the sequence by the domain: 61.00 %

HMM information View help on HMM parameters

HMM build commands:
build method: hmmbuild -o /dev/null HMM SEED
search method: hmmsearch -Z 26740544 -E 1000 --cpu 4 HMM pfamseq
Model details:
Parameter Sequence Domain
Gathering cut-off 25.0 25.0
Trusted cut-off 70.2 69.7
Noise cut-off 20.7 19.5
Model length: 58
Family (HMM) version: 1
Download: download the raw HMM for this family

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