Solubilization of hydrophobic peptides by reversible cysteine PEGylation

Biochem Biophys Res Commun. 1998 Apr 17;245(2):618-21. doi: 10.1006/bbrc.1998.8493.

Abstract

"PEG-a-Cys" reagent, synthesized by the esterification of monomethoxy-poly(ethylene glycol) (avg. MW = 5 kDa) to Ellman's reagent [5,5'-dithiobis(2-nitrobenzoic acid)], is shown to "PEGylate" reversibly the cysteine residue of a 25-residue synthetic hydrophobic peptide (H2N-REAAALAAAAALAAWAALCPARRRR-CO2H) designed to model a transmembrane segment of a membrane protein. A mixed disulfide bond was formed between the reagent and the peptide that was readily cleaved with the mild reducing agent tricarboxyethylphosphine hydrochloride (TCEP.HCl). Carboxypeptidase B digestion of the charged carboxyl terminus of the peptide through to the Ala residue--which mimics the enzymatic cleavage of a TM segment from a fusion protein--releases a highly hydrophobic peptide. A time-dependent decrease in the amplitude of the digested peptide circular dichroism (CD) spectra was attributed to the aggregation and/or precipitation of the peptide. While PEGylation of the peptide with PEG-a-Cys had a negligible effect on conformation, it inhibited the loss of CD amplitude in both intact and digested peptides, suggesting that it was effective in solubilization of hydrophobic peptides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Carboxypeptidase B
  • Carboxypeptidases / metabolism
  • Circular Dichroism
  • Cysteine / chemistry*
  • Disulfides / metabolism
  • Dithionitrobenzoic Acid / analogs & derivatives
  • Dithionitrobenzoic Acid / metabolism
  • Molecular Conformation
  • Molecular Sequence Data
  • Molecular Structure
  • Peptides / chemistry*
  • Phosphines / metabolism
  • Polyethylene Glycols / chemistry
  • Solubility*

Substances

  • Disulfides
  • Peptides
  • Phosphines
  • tris(2-carboxyethyl)phosphine
  • Polyethylene Glycols
  • Dithionitrobenzoic Acid
  • Carboxypeptidases
  • Carboxypeptidase B
  • Cysteine