title: Widening and diversifying the proteome capture by combinatorial peptide ligand libraries via Alcian Blue dye binding.
abstract: Combinatorial peptide ligand libraries (CPLLs) tend to bind complex molecules such as dyes due to their aromatic, heterocyclic, hydrophobic, and ionic nature that may affect the protein capture specificity. In this experimental work Alcian Blue 8GX, a positively charged phthalocyanine dye well-known to bind to glycoproteins and to glucosaminoglycans, was adsorbed on a chemically modified CPLL solid phase, and the behavior of the resulting conjugate was then investigated. The control and dye-adsorbed beads were used to harvest the human urinary proteome at physiological pH, this resulting in a grand total of 1151 gene products identified after the capture. Although the Alcian Blue-modified CPLL incremented the total protein capture by 115 species, it particularly enriched some families among the harvested proteins, such as glycoproteins and nucleotide-binding proteins. This study teaches that it is possible, via the two combined harvest mechanisms, to drive the CPLL capture toward the enrichment of specific protein categories.
authors: †Nephrology, Dialysis, Transplantation Unit and Laboratory on Pathophysiology of Uremia, Istituto Giannina Gaslini, Genoa 16148, Italy.
authors: Giovanni
authors: G
authors: Candiano
authors: †Nephrology, Dialysis, Transplantation Unit and Laboratory on Pathophysiology of Uremia, Istituto Giannina Gaslini, Genoa 16148, Italy.
authors: Laura
authors: L
authors: Santucci
authors: ‡Core Facilities-Proteomics Laboratory, Istituto Giannina Gaslini, Genoa 16148, Italy.
authors: Andrea
authors: A
authors: Petretto
authors: ‡Core Facilities-Proteomics Laboratory, Istituto Giannina Gaslini, Genoa 16148, Italy.
authors: Chiara
authors: C
authors: Lavarello
authors: ‡Core Facilities-Proteomics Laboratory, Istituto Giannina Gaslini, Genoa 16148, Italy.
authors: Elvira
authors: E
authors: Inglese
authors: †Nephrology, Dialysis, Transplantation Unit and Laboratory on Pathophysiology of Uremia, Istituto Giannina Gaslini, Genoa 16148, Italy.
authors: Maurizio
authors: M
authors: Bruschi
authors: †Nephrology, Dialysis, Transplantation Unit and Laboratory on Pathophysiology of Uremia, Istituto Giannina Gaslini, Genoa 16148, Italy.
authors: Gian Marco
authors: GM
authors: Ghiggeri
authors: §EB JAM-Conseil, 92200 Neuilly sur-Seine, Paris, France.
authors: Egisto
authors: E
authors: Boschetti
authors: ∥Department of Chemistry, Materials and Chemical Engineering, "Giulio Natta", Politecnico di Milano, Via Mancinelli 7, Milano 20131, Italy.
authors: Pier Giorgio
authors: PG
authors: Righetti
mesh_terms: D000327:Adsorption; D000328:Adult; D000423:Alcian Blue / Q000737:chemistry*; D001665:Binding Sites; D005260:Female; D064368:Healthy Volunteers; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008024:Ligands; D008297:Male; D008875:Middle Aged; D015394:Molecular Structure; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D020543:Proteome / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D013499:Surface Properties; D016482:Urinalysis / Q000379:methods*
chemical_list: D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D020543:Proteome; D000423:Alcian Blue