DNA sequence-directed assembly of two peptide bioconjugates.
Peptide bioconjugates combine the molecular recognition features of the polypeptide from a DNA-binding protein and the DNA-sensitive fluorescence of an intercalating dye. Here, DNA template-directed assembly of two bioconjugate probes was examined by steady-state fluorescence resonance energy transfer and time-resolved single photon counting. The Förster critical distance was determined to be approximately 26 A for the oxazole yellow and thiazole orange donor-acceptor pair. The efficiency of energy transfer for two bioconjugates was a function of the number of intervening base pairs between two DNA cognate sites. These probes were sufficiently sensitive to detect sequence dependent curvature and polypeptide induced bending of the DNA. Molecular probes capable of examining spatial aspects of protein complexes at promoter sites could yield important information about the early events in transcription initiation.
Thompson
M
M
Department of Chemistry, Michigan Technological University, Houghton, MI 49931, USA. mthomps@mtu.edu
D000595:Amino Acid Sequence; D001483:Base Sequence; D052160:Benzothiazoles / Q000737:chemistry; D001583:Benzoxazoles / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D004254:DNA Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D007364:Intercalating Agents / Q000138:chemical synthesis / Q000737:chemistry*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D011804:Quinolines / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D019895:Transposases / Q000737:chemistry / Q000235:genetics
D052160:Benzothiazoles; D001583:Benzoxazoles; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D010446:Peptide Fragments; D010455:Peptides; D011804:Quinolines; C097015:oxazole yellow; C051000:thiazole orange; D004247:DNA; D004254:DNA Nucleotidyltransferases; C059997:Hin recombinase; D019895:Transposases