ProP-PD/PDZ-Seq
Proteomic Peptide-Phage Display / PDZ Domains Sequencing
ProP-PD is a protocol that identifies short linear motif (SLiM) interactions or PDZ domains (PDZ-Seq) (Ivarsson et al., 2014) (Ernst et al., 2010). ProP-PD can be used for the direct identification of ligands with potential biological relevance. This method contrasts with the commonly used combinatorial phage display, where highly diverse libraries are used to establish preferred binding motifs.
In this method, C-terminal sequences are first identified and an oligonucleotide library is created. Next, a phage display library is constructed, which is hybridized to immobilized bait proteins. After multiple rounds of selection, the selected pools that bind to the PDZ domains are sequenced and counted. Deep sequencing provides detailed information pertaining to the motifs that interact with specific proteins.
Advantages:
- Direct identification of the ligands
Disadvantages:
- Construction of the oligonucleotide library may be biased
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
None available yet
References:
Garrido-Urbani S., Garg P., Ghossoub R., et al. Proteomic peptide phage display uncovers novel interactions of the PDZ1-2 supramodule of syntenin. FEBS Lett. 2016;590:3-12
Ivarsson Y., Arnold R., McLaughlin M., et al. Large-scale interaction profiling of PDZ domains through proteomic peptide-phage display using human and viral phage peptidomes. Proc Natl Acad Sci U S A. 2014;111:2542-2547
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History: ProP-PD/PDZ-Seq
Revision by sbrumpton on 2017-06-21 07:50:21 - Show/Hide
Proteomic Peptide-Phage Display / PDZ Domains Sequencing
ProP-PD is a protocol that identifies short linear motif (SLiM) interactions or PDZ domains (PDZ-Seq) (Ivarsson et al., 2014) (Ernst et al., 2010). ProP-PD can be used for the direct identification of ligands with potential biological relevance. This method contrasts with the commonly used combinatorial phage display, where highly diverse libraries are used to establish preferred binding motifs.
In this method, C-terminal sequences are first identified and an oligonucleotide library is created. Next, a phage display library is constructed, which is hybridized to immobilized bait proteins. After multiple rounds of selection, the selected pools that bind to the PDZ domains are sequenced and counted. Deep sequencing provides detailed information pertaining to the motifs that interact with specific proteins.
Advantages:- Direct identification of the ligands
Disadvantages:- Construction of the oligonucleotide library may be biased
Reagents:Illumina Library prep and Array Kit SelectorReviews:None available yet
References:Garrido-Urbani S., Garg P., Ghossoub R., et al. Proteomic peptide phage display uncovers novel interactions of the PDZ1-2 supramodule of syntenin. FEBS Lett. 2016;590:3-12Ivarsson Y., Arnold R., McLaughlin M., et al. Large-scale interaction profiling of PDZ domains through proteomic peptide-phage display using human and viral phage peptidomes. Proc Natl Acad Sci U S A. 2014;111:2542-2547