On-the-resin N-terminal modification of long synthetic peptides

Anal Biochem. 2012 May 15;424(2):137-9. doi: 10.1016/j.ab.2012.02.032. Epub 2012 Mar 1.

Abstract

Here we present a highly efficient protocol for on-the-resin coupling of fluorescent dyes or other functional groups to the N-termini of synthetic peptides prior to cleavage and deprotection. The protocol avoids expensive preactivated dyes and instead employs carboxylated dyes activated by large amounts of coupling reagents. The protocol was used to label peptides with low reactivity such as long hydrophobic peptides and peptides with strong tendencies to form sterically shielding structures or aggregates in solution. In all cases, the yields far exceeded those from commercially available preactivated compounds.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Ethylamines / chemistry
  • Fluorescent Dyes
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Immobilized Proteins / analysis
  • Immobilized Proteins / chemistry
  • Molecular Sequence Data
  • Organophosphorus Compounds / chemistry
  • Peptides / analysis
  • Peptides / chemistry*
  • Receptor, Fibroblast Growth Factor, Type 1 / analysis
  • Receptor, Fibroblast Growth Factor, Type 1 / chemistry*
  • Receptor, Fibroblast Growth Factor, Type 2 / analysis
  • Receptor, Fibroblast Growth Factor, Type 2 / chemistry*
  • Staining and Labeling / methods*

Substances

  • Ethylamines
  • Fluorescent Dyes
  • Immobilized Proteins
  • Organophosphorus Compounds
  • Peptides
  • N,N-diisopropylethylamine
  • FGFR1 protein, human
  • FGFR2 protein, human
  • Receptor, Fibroblast Growth Factor, Type 1
  • Receptor, Fibroblast Growth Factor, Type 2