SFG analysis of surface bound proteins: a route towards structure determination

Tobias Weidner, David G. Castner

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Abstract

The surface of a material is rapidly covered with proteins once that material is placed in a biological environment. The structure and function of these bound proteins play a key role in the interactions and communications of the material with the biological environment. Thus, it is crucial to gain a molecular level understanding of surface bound protein structure. While X-ray diffraction and solution phase NMR methods are well established for determining the structure of proteins in the crystalline or solution phase, there is not a corresponding single technique that can provide the same level of structural detail about proteins at surfaces or interfaces. However, recent advances in sum frequency generation (SFG) vibrational spectroscopy have significantly increased our ability to obtain structural information about surface bound proteins and peptides. A multi-technique approach of combining SFG with (1) protein engineering methods to selectively introduce mutations and isotopic labels, (2) other experimental methods such as time-of-flight secondary ion mass spectrometry (ToF-SIMS) and near edge X-ray absorption fine structure (NEXAFS) to provide complementary information, and (3) molecular dynamic (MD) simulations to extend the molecular level experimental results is a particularly promising route for structural characterization of surface bound proteins and peptides. By using model peptides and small proteins with well-defined structures, methods have been developed to determine the orientation of both backbone and side chains to the surface.

Original languageEnglish
JournalPhysical chemistry chemical physics : PCCP
Volume15
Issue30
Pages (from-to)12516-24
Number of pages9
DOIs
Publication statusPublished - 2013
Externally publishedYes

Keywords

  • Isotope Labeling
  • Molecular Dynamics Simulation
  • Peptides
  • Polystyrenes
  • Protein Structure, Secondary
  • Proteins
  • Spectrometry, Mass, Secondary Ion
  • Surface Properties
  • X-Ray Absorption Spectroscopy
  • Journal Article
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

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