scholarly journals Large-scale interaction profiling of PDZ domains through proteomic peptide-phage display using human and viral phage peptidomes

2014 ◽  
Vol 111 (7) ◽  
pp. 2542-2547 ◽  
Author(s):  
Y. Ivarsson ◽  
R. Arnold ◽  
M. McLaughlin ◽  
S. Nim ◽  
R. Joshi ◽  
...  
2017 ◽  
Author(s):  
Gustav N. Sundell ◽  
Roland Arnold ◽  
Muhammad Ali ◽  
Julien Orts ◽  
Peter Güntert ◽  
...  

We report phosphomimetic proteomic peptide-phage display, a powerful large-scale method for finding ligands of short linear motif binding domains that simultaneously pinpoint functional Ser/Thr phosphosites in three steps. First, we computationally designed an oligonucleotide library encoding all human C-terminal peptides containing known or predicted Ser/Thr phosphosites and phosphomimetic variants thereof. Second, we incorporated these oligonucleotides into a phage library. Third, we screened the six PDZ (PSD-95/Dlg/ZO-1) domains of Scribble and DLG1 for binding and identified known and novel ligands from the human proteome, and whether these interactions may be regulated by ligand phosphorylation. We demonstrate that the Scribble PDZ domains preferentially bind to ligands with phosphomimetic mutations at two distinct positions, and show that the equilibrium dissociation constant for Scribble PDZ1 with the C-terminal peptide of RPS6KA2 is enhanced over four-fold by phosphorylation. We elucidate the molecular determinants of phosphopeptide binding through NMR structure determination and mutational analysis. Finally, we discuss the role of Ser/Thr phosphorylation as a switching mechanism of PDZ domain interactions.


2000 ◽  
Vol 165 (7) ◽  
pp. 3830-3838 ◽  
Author(s):  
Mark A. Myers ◽  
Janet M. Davies ◽  
Jonathan C. Tong ◽  
James Whisstock ◽  
Marita Scealy ◽  
...  

2011 ◽  
Vol 181 (2-4) ◽  
pp. 291-300 ◽  
Author(s):  
Renan O. Clara ◽  
Tatiane S. Soares ◽  
Ricardo J.S. Torquato ◽  
Cássia A. Lima ◽  
Renata O.M. Watanabe ◽  
...  

Author(s):  
Paul Giguere ◽  
Scott W. Formica ◽  
Wayne M. Harding ◽  
Michele R. Cummins

Designing online trainings or courses for large numbers of participants can prove to be challenging for instructors and facilitators. Online learning environments need to be structured in a way that preserves actual or perceived levels of interaction, participant perceptions of value and utility, and achievement of the learning objectives. This chapter describes five Large-Scale Interaction Strategies that offer guidance for addressing some of these online instructional design issues. Evaluation data are presented in support of two of the strategies, and recommendations are provided about how future research in this area might be conducted.


2020 ◽  
Vol 48 (W1) ◽  
pp. W200-W207
Author(s):  
Simone Puccio ◽  
Giorgio Grillo ◽  
Arianna Consiglio ◽  
Maria Felicia Soluri ◽  
Daniele Sblattero ◽  
...  

Abstract High-Throughput Sequencing technologies are transforming many research fields, including the analysis of phage display libraries. The phage display technology coupled with deep sequencing was introduced more than a decade ago and holds the potential to circumvent the traditional laborious picking and testing of individual phage rescued clones. However, from a bioinformatics point of view, the analysis of this kind of data was always performed by adapting tools designed for other purposes, thus not considering the noise background typical of the ‘interactome sequencing’ approach and the heterogeneity of the data. InteractomeSeq is a web server allowing data analysis of protein domains (‘domainome’) or epitopes (‘epitome’) from either Eukaryotic or Prokaryotic genomic phage libraries generated and selected by following an Interactome sequencing approach. InteractomeSeq allows users to upload raw sequencing data and to obtain an accurate characterization of domainome/epitome profiles after setting the parameters required to tune the analysis. The release of this tool is relevant for the scientific and clinical community, because InteractomeSeq will fill an existing gap in the field of large-scale biomarkers profiling, reverse vaccinology, and structural/functional studies, thus contributing essential information for gene annotation or antigen identification. InteractomeSeq is freely available at https://InteractomeSeq.ba.itb.cnr.it/


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