Gold-ATP.
The synthesis, purification, and chemical analysis of two covalent conjugates between ATP and undecagold are described, one in which gold is attached to the ribose moiety of ATP and the other in which it is attached to the N-6 position of the adenine base. The former probe was then used to bind to two ATP binding proteins, the helicase DnaB and the chaperone DnaK. After purification from unbound gold by column chromatography, binding was measured by UV-Vis spectroscopy, then the protein and gold were visualized by scanning transmission electron microscopy. Binding was observed with the conjugates, and virtually no binding occurred in the control of undecagold without the ATP attached. This new probe may be useful for studying nucleotide binding sites on proteins or for labeling nucleic acids or oligonucleotides directly.
Hainfeld
J F
JF
Biology Department, Brookhaven National Laboratory, Upton, New York 11973, USA.
Liu
W
W
Barcena
M
M
D000255:Adenosine Triphosphate / Q000032:analysis* / Q000737:chemistry; D000818:Animals; D001426:Bacterial Proteins; D001665:Binding Sites; D004265:DNA Helicases / Q000032:analysis; D053488:DnaB Helicases; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D006046:Gold / Q000032:analysis / Q000737:chemistry; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis / Q000378:metabolism; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D017348:Microscopy, Electron, Scanning Transmission; D018832:Molecular Chaperones; D015335:Molecular Probes / Q000032:analysis / Q000138:chemical synthesis / Q000302:isolation & purification; D050607:Organogold Compounds; D009942:Organometallic Compounds / Q000737:chemistry*; D010759:Phosphorus Isotopes / Q000032:analysis; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D008024:Ligands; D018832:Molecular Chaperones; D015335:Molecular Probes; D050607:Organogold Compounds; D009942:Organometallic Compounds; D010759:Phosphorus Isotopes; C035797:undecagold; D006046:Gold; D000255:Adenosine Triphosphate; C063842:dnaK protein, E coli; D004265:DNA Helicases; D053488:DnaB Helicases