Screening of Yeast Display Libraries of Enzymatically Treated Peptides to Discover Macrocyclic Peptide Ligands.
Adaptor Proteins, Signal Transducing
/ chemistry
Albumins
/ metabolism
Binding Sites
Combinatorial Chemistry Techniques
Endoplasmic Reticulum
/ metabolism
Flow Cytometry
Ligands
Muramidase
/ metabolism
Peptides, Cyclic
/ isolation & purification
Protein Binding
Protein Engineering
/ methods
Transcription Factors
/ chemistry
Transglutaminases
/ metabolism
YAP-Signaling Proteins
Yeasts
/ genetics
WW domain
Yes Associated Protein (YAP) 65
cyclic peptide
transglutaminase
yeast display library
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
05 Feb 2021
05 Feb 2021
Historique:
received:
23
11
2020
revised:
12
01
2021
accepted:
26
01
2021
entrez:
10
2
2021
pubmed:
11
2
2021
medline:
3
9
2021
Statut:
epublish
Résumé
We present the construction and screening of yeast display libraries of post-translationally modified peptides wherein site-selective enzymatic treatment of linear peptides is achieved using bacterial transglutaminase. To this end, we developed two alternative routes, namely (i) yeast display of linear peptides followed by treatment with recombinant transglutaminase in solution; or (ii) intracellular co-expression of linear peptides and transglutaminase to achieve peptide modification in the endoplasmic reticulum prior to yeast surface display. The efficiency of peptide modification was evaluated via orthogonal detection of epitope tags integrated in the yeast-displayed peptides by flow cytometry, and via comparative cleavage of putative cyclic vs. linear peptides by tobacco etch virus (TEV) protease. Subsequently, yeast display libraries of transglutaminase-treated peptides were screened to isolate binders to the N-terminal region of the Yes-Associated Protein (YAP) and its WW domains using magnetic selection and fluorescence activated cell sorting (FACS). The identified peptide cyclo[
Identifiants
pubmed: 33562883
pii: ijms22041634
doi: 10.3390/ijms22041634
pmc: PMC7915732
pii:
doi:
Substances chimiques
Adaptor Proteins, Signal Transducing
0
Albumins
0
Ligands
0
Peptides, Cyclic
0
Transcription Factors
0
YAP-Signaling Proteins
0
YAP1 protein, human
0
Transglutaminases
EC 2.3.2.13
Muramidase
EC 3.2.1.17
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : National Science Foundation
ID : CBET 1510845
Organisme : National Institute of Health
ID : NIH T32 GM008776
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