title: Electrostatic interactions observed when imaging proteins with the atomic force microscope.
abstract: The atomic force microscope (AFM) is now an established and valuable tool for the study of biological macromolecules in aqueous environments. In this paper we form a patterned boundary via the microcontact printing of individually isolated proteins, covalently attached to a solid support. We use this boundary to investigate electrostatic interactions that can occur between an AFM tip and a protein surface during imaging in solution. The observed height variations of the protein film are found to be a combination of not only structural considerations and thickness of the protein film, but also the repulsive contribution from electrostatic interactions between the AFM tip and the sample. These variations in measured heights of the protein surface can be described by Derjaguin, Landau, Verway, Overbeek (DLVO) theory. Our experimental results show that height measurements can be manipulated either negatively or positively by adjusting the pH and concentration of the electrolyte buffer that is utilised.
authors.affiliation: Laboratory of Biophysics and Surface Analysis, School of Pharmaceutical Sciences, University of Nottingham, University Park, Nottingham NG7 2RD, UK.
authors.forename: J P
authors.initials: JP
authors.lastname: Rossell
authors.forename: S
authors.initials: S
authors.lastname: Allen
authors.forename: M C
authors.initials: MC
authors.lastname: Davies
authors.forename: C J
authors.initials: CJ
authors.lastname: Roberts
authors.forename: S J B
authors.initials: SJ
authors.lastname: Tendler
authors.forename: P M
authors.initials: PM
authors.lastname: Williams
mesh_terms: D000465:Algorithms; D002021:Buffers; D002374:Catalase / Q000648:ultrastructure; D004573:Electrolytes / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D018625:Microscopy, Atomic Force / Q000295:instrumentation*; D011506:Proteins / Q000648:ultrastructure*; D055672:Static Electricity; D014867:Water / Q000737:chemistry
chemical_list: D002021:Buffers; D004573:Electrolytes; D011506:Proteins; D014867:Water; D006046:Gold; D002374:Catalase