Plucking, pillaging and plundering proteomes with combinatorial peptide ligand libraries

Pier Giorgio Righetti, Egisto Boschetti, Alberto Zanella, Elisa Fasoli, Attilio Citterio

Research output: Contribution to journalArticle

Abstract

Recent developments in the technique of combinatorial peptide ligand libraries, for enhancing the visibility of the low-abundance proteome, are reviewed here. Novel en bloc elution systems, allowing essentially complete proteome recovery in a single step, are reported here, particularly, en bloc elution with 3-5% boiling sodium dodecyl sulphate (SDS) or in urea-thiourea-CHAPS added with either 40 mM formic acid or 25 mM cysteic acid. Novel capturing systems are also discussed: in particular, although capturing at pH 7.2 in physiological saline has always been recommended, it is shown that capturing also at acidic (pH 3.8) and alkaline (pH 9.5) values substantially increments the total captured protein population. Some examples of detection of novel proteins by the described methodology are also discussed. In particular, in the case of venom proteins, where essentially all components had been detected and fully described by conventional means, the application of the ligand library technology allowed the discovery of two, previously unreported, trace enzymes necessary for the maintenance of the native structure of venom components, namely peroxiredoxin and glutaminyl cyclase. In the case of the red blood cell (RBC) cytoplasmic proteome, where a grand total of 1570 components of the 2% minority proteomes have been identified, these findings allowed to unravel the genetic defect of a rare RBC disease, called congenital dyserythropoietic anemia type II. The mutations are located in the SEC23B gene coding for the SEC23B protein, detected for the first time in the RBC proteome thanks to the peptide capturing technology.

Original languageEnglish
Pages (from-to)893-900
Number of pages8
JournalJournal of Chromatography A
Volume1217
Issue number6
DOIs
Publication statusPublished - Feb 5 2010

Keywords

  • Elution systems
  • Ligand library
  • Low-abundance proteome
  • Peptide ligands

ASJC Scopus subject areas

  • Analytical Chemistry
  • Organic Chemistry
  • Biochemistry

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