CUED Publications database

Mimicking nature: Phosphopeptide enrichment using combinatorial libraries of affinity ligands.

Batalha, IL and Zhou, H and Lilley, K and Lowe, CR and Roque, ACA (2016) Mimicking nature: Phosphopeptide enrichment using combinatorial libraries of affinity ligands. J Chromatogr A, 1457. pp. 76-87.

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Abstract

Phosphorylation is a reversible post-translational modification of proteins that controls a plethora of cellular processes and triggers specific physiological responses, for which there is a need to develop tools to characterize phosphorylated targets efficiently. Here, a combinatorial library of triazine-based synthetic ligands comprising 64 small molecules has been rationally designed, synthesized and screened for the enrichment of phosphorylated peptides. The lead candidate (coined A8A3), composed of histidine and phenylalanine mimetic components, showed high binding capacity and selectivity for binding mono- and multi-phosphorylated peptides at pH 3. Ligand A8A3 was coupled onto both cross-linked agarose and magnetic nanoparticles, presenting higher binding capacities (100-fold higher) when immobilized on the magnetic support. The magnetic adsorbent was further screened against a tryptic digest of two phosphorylated proteins (α- and β-caseins) and one non-phosphorylated protein (bovine serum albumin, BSA). The MALDI-TOF mass spectra of the eluted peptides allowed the identification of nine phosphopeptides, comprising both mono- and multi-phosphorylated peptides.

Item Type: Article
Uncontrolled Keywords: Affinity Iron oxide Magnetic nanoparticles Phosphopeptide enrichment Phosphoproteomics Synthetic ligands Adsorption Animals Biomimetic Materials Caseins Cattle Combinatorial Chemistry Techniques Ferric Compounds Ligands Nanoparticles Phosphopeptides Phosphorylation Serum Albumin, Bovine Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Triazines
Subjects: UNSPECIFIED
Divisions: Div B > Solid State Electronics and Nanoscale Science
Depositing User: Cron Job
Date Deposited: 17 Jul 2017 19:13
Last Modified: 18 Nov 2017 21:48
DOI: