Use of engineered unique cysteine residues to facilitate oriented coupling of proteins directly to a gold substrate

Photochem Photobiol. 2011 Sep-Oct;87(5):1050-7. doi: 10.1111/j.1751-1097.2011.00948.x. Epub 2011 Jul 28.

Abstract

A prerequisite for any "lab on a chip" device that utilizes an electrical signal from the sensor protein is the ability to attach the protein in a specific orientation onto a conducting substrate. Here, we demonstrate the covalent attachment to a gold surface of light-harvesting membrane proteins, from Rhodobacter sphaeroides, via cysteine (Cys) residues engineered on either the cytoplasmic or periplasmic face. This simple directed attachment is superior in its ability to retain light-harvesting complex (LHC) function, when compared to a similar attachment procedure utilizing a self-assembled monolayer on gold. LH 1 has previously been observed to have superior photostability over LH 2 (Magis et al. [2010] Biochim. Biophys. Acta, 1798, 637-645); this characteristic is maintained even with the introduction of Cys residues.

Publication types

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

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Cysteine / chemistry*
  • Cysteine / metabolism
  • Gold / chemistry
  • Lab-On-A-Chip Devices*
  • Light
  • Light-Harvesting Protein Complexes / chemistry*
  • Light-Harvesting Protein Complexes / genetics
  • Light-Harvesting Protein Complexes / metabolism
  • Membrane Proteins / chemistry*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Microscopy, Atomic Force
  • Mutagenesis, Site-Directed
  • Photochemical Processes / radiation effects
  • Protein Binding
  • Rhodobacter sphaeroides / chemistry*

Substances

  • Bacterial Proteins
  • Light-Harvesting Protein Complexes
  • Membrane Proteins
  • Gold
  • Cysteine