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Screening and Identification of PLK1-Polo Box Binding Peptides by Highthroughput Sequencing of Phage-selected Libraries

Author(s):

Nousheen Bibi, Hafsa Niaz, Ted Hupp, Mohammad Amjad Kamal and Sajid Rashid*   Pages 1 - 13 ( 13 )

Abstract:


Background: Human proteome contains a plethora of short linear peptide motifs that is crucial for signaling and other cellular processes. These motifs are difficult to identify due to lack of systematic approach for their detection.

Objectives: Here we demonstrate the use of peptide phage display in combination with high throughput next generation sequencing to identify enriched peptide sequences through biopanning process against polo box domain (PBD) of mitotic polo like kinase 1 (Plk1).

Method: Purified recombinant Plk1 and two unrelated controls namely B-lymphocyte antigen (CD20) and fluorescent protein (mCherry) were subjected to peptide phage display analysis. Bacterially- propagated phage DNA was amplified by PCR using triplet bar coded primers to tag the pool from each amplicon.

Results: Proteomic peptide phage display along with next generation sequencing and Bioinformatics analysis demonstrated several known and putative novel interactions which were potentially related to Plk1-PBD. With our strategy, we were able to identify and characterize several Plk1-PBD binding peptides, as well as to define more precisely consensus sequences.

Conclusion: We believe that this information could provide valuable tool for exploring novel interaction involved in Plk1 signaling as well as to choose peptides for Plk1 specific drug development.

Keywords:

Plk1 polo box domain, peptide phage display, next generation sequencing, consensus binding motif.

Affiliation:

Shaheed Benazir Bhutto Women University, Peshawar, National Center for Bioinformatics, Quaid-i-Azam University, Islamabad, Edinburgh Cancer Research Center, University of Edinburgh, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, National Center for Bioinformatics, Quaid-i-Azam University, Islamabad



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