Scanning force microscopy of nucleic acid complexes.
SFM is a viable effective method for determining the mode of binding, the extent of binding, and the site of binding of intercalators to nucleic acids. Establishing the presence of a groove-bound ligand can be achieved either by competitive binding experiments with a well-defined intercalator (minor groove) or by changes in apparent contrast (major groove). In our opinion, SFM has an important role in resolving the structural polymorphisms for small molecule-DNA complexes. Application of these assays in the study of polyintercalator molecules is currently underway in our laboratory. SFM is an important, new tool in the study of protein-DNA complexes. New insights into the structure and function of these complexes are enabled by real-time visualization. Currently the temporal resolution of the SFM limits the degree to which definitive rate data can be determined. Several binding and unbinding events could take place in the time it takes to acquire one image. New developments in SFM technology will allow faster scanning and will improve the temporal resolution of so-called SFM movies. To this end, the Hansma group is developing small cantilevers and improved optical deflection systems to enable intermittent imaging at scanning rates of 1.7 sec per image. These improvements will enable SFM visualization of complex biological processes as they occur, one molecule at a time.
Lillehei
P T
PT
School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
Bottomley
L A
LA
D001665:Binding Sites; D004247:DNA / Q000737:chemistry / Q000648:ultrastructure; D066298:In Vitro Techniques; D007364:Intercalating Agents / Q000737:chemistry; D046911:Macromolecular Substances; D018625:Microscopy, Atomic Force / Q000295:instrumentation / Q000379:methods*; D009696:Nucleic Acids / Q000737:chemistry* / Q000648:ultrastructure*; D004364:Pharmaceutical Preparations / Q000737:chemistry; D011506:Proteins / Q000737:chemistry / Q000648:ultrastructure
D007364:Intercalating Agents; D046911:Macromolecular Substances; D009696:Nucleic Acids; D004364:Pharmaceutical Preparations; D011506:Proteins; D004247:DNA