Comparison of Zirconium Phosphonate-Modified Surfaces for Immobilizing Phosphopeptides and Phosphate-Tagged Proteins

Langmuir. 2016 Jun 7;32(22):5480-90. doi: 10.1021/acs.langmuir.6b01020. Epub 2016 May 23.

Abstract

Different routes for preparing zirconium phosphonate-modified surfaces for immobilizing biomolecular probes are compared. Two chemical-modification approaches were explored to form self-assembled monolayers on commercially available primary amine-functionalized slides, and the resulting surfaces were compared to well-characterized zirconium phosphonate monolayer-modified supports prepared using Langmuir-Blodgett methods. When using POCl3 as the amine phosphorylating agent followed by treatment with zirconyl chloride, the result was not a zirconium-phosphonate monolayer, as commonly assumed in the literature, but rather the process gives adsorbed zirconium oxide/hydroxide species and to a lower extent adsorbed zirconium phosphate and/or phosphonate. Reactions giving rise to these products were modeled in homogeneous-phase studies. Nevertheless, each of the three modified surfaces effectively immobilized phosphopeptides and phosphopeptide tags fused to an affinity protein. Unexpectedly, the zirconium oxide/hydroxide modified surface, formed by treating the amine-coated slides with POCl3/Zr(4+), afforded better immobilization of the peptides and proteins and efficient capture of their targets.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Archaeal Proteins / chemistry*
  • DNA-Binding Proteins / chemistry*
  • Organophosphonates / chemistry*
  • Phosphopeptides / chemistry*
  • Phosphoproteins / chemistry*
  • Sulfolobus acidocaldarius / chemistry*
  • Surface Properties
  • Zirconium / chemistry*

Substances

  • Archaeal Proteins
  • DNA-Binding Proteins
  • Organophosphonates
  • Phosphopeptides
  • Phosphoproteins
  • Sac7 protein, Sulfolobus
  • Zirconium