Electrodeposition procedure of E. coli RNA polymerase onto gold and deposition of E. coli RNA polymerase onto mica for observation with scanning force microscopy.
Molecules of the transcriptional enzyme E. coli RNA polymerase (RNAP) have been deposited using three different deposition methods: (1) passive adsorption onto gold, (2) electrochemical adsorption onto gold and (3) adsorption onto mica. In all cases SFM imaging was straightforward and reliable, and surface coverage by the protein varied with deposition conditions as expected. To determine the nature of the electrochemical treatment on the gold substrate, cyclic voltammetry was performed with various chemical solutions. Finally, a comparison is made between the SFM images of RNAP obtained with these methods and STM images obtained earlier. Both STM and SFM show strikingly similar results; however, heights and widths of individual molecules differ.
Keller
R W
RW
Department of Chemistry, University of New Mexico, Albuquerque 87131.
Keller
D J
DJ
Bear
D
D
Vasenka
J
J
Bustamante
C
C
D000327:Adsorption; D000538:Aluminum Silicates; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000648:ultrastructure*; D004563:Electrochemistry; D004926:Escherichia coli / Q000201:enzymology*; D006046:Gold; D008853:Microscopy / Q000379:methods*; D013048:Specimen Handling
D000538:Aluminum Silicates; D006046:Gold; D012321:DNA-Directed RNA Polymerases; C011934:mica