Single molecule studies of protein folding using atomic force microscopy.
Atomic force microscopy (AFM) offers new insights into the ability of proteins to resist mechanical force. The technique has been opened up by the availability of easy-to-use instruments that are commercially available, so that the technique no longer relies on the need to build instruments in the lab. Indeed it may become common for AFM instruments to sit beside stopped-flow apparatus in protein folding laboratories. In this chapter, we describe the instrument set-up, the preparation of suitable protein substrate, and the collection of data. Data selection and analysis are more complex than for conventional stopped-flow ensemble studies, but offer new insights into the function of proteins in vivo.
Ng
Sean P
SP
Department of Chemistry, University of Cambridge, Medical Research Council Center for Protein Engineering, Cambridge, UK.
Randles
Lucy G
LG
Clarke
Jane
J
D018625:Microscopy, Atomic Force / Q000295:instrumentation / Q000379:methods*; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*
D011506:Proteins