Ultraviolet resonance Raman study of drug binding in dihydrofolate reductase, gyrase, and catechol O-methyltransferase.
This paper presents a study of the use of ultraviolet resonance Raman (UVRR) spectroscopic methods as a means of elucidating aspects of drug-protein interactions. Some of the RR vibrational bands of the aromatic amino acids tyrosine and tryptophan are sensitive to the microenvironment, and the use of UV excitation radiation allows selective enhancement of the spectral features of the aromatic amino acids, enabling observation specifically of their change in microenvironment upon drug binding. The three drug-protein systems investigated in this study are dihydrofolate reductase with its inhibitor trimethoprim, gyrase with novobiocin, and catechol O-methyltransferase with dinitrocatechol. It is demonstrated that UVRR spectroscopy has adequate sensitivity to be a useful means of detecting drug-protein interactions in those systems for which the electronic absorption of the aromatic amino acids changes because of hydrogen bonding and/or possible dipole-dipole and dipole-polarizability interactions with the ligand.
Couling
V W
VW
Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, England.
Fischer
P
P
Klenerman
D
D
Huber
W
W
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002394:Catechol O-Methyltransferase / Q000737:chemistry* / Q000378:metabolism; D002396:Catechols / Q000378:metabolism; D004250:DNA Topoisomerases, Type II / Q000737:chemistry* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D004926:Escherichia coli; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009675:Novobiocin / Q000378:metabolism*; D017354:Point Mutation; D011487:Protein Conformation; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013059:Spectrum Analysis, Raman / Q000379:methods; D013211:Staphylococcus aureus / Q000201:enzymology; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D014295:Trimethoprim / Q000378:metabolism*; D014364:Tryptophan; D014443:Tyrosine; D014466:Ultraviolet Rays
D002396:Catechols; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D009675:Novobiocin; D014443:Tyrosine; D014364:Tryptophan; D014295:Trimethoprim; D013762:Tetrahydrofolate Dehydrogenase; D002394:Catechol O-Methyltransferase; D004250:DNA Topoisomerases, Type II; C057726:OR486