title: Spectroscopic characterization of furosemide binding to human carbonic anhydrase II.
abstract: This study reports the interaction between furosemide and human carbonic anhydrase II (hCA II) using fluorescence, UV-vis and circular dichroism (CD) spectroscopy. Fluorescence data indicated that furosemide quenches the intrinsic fluorescence of the enzyme via a static mechanism and hydrogen bonding and van der Walls interactions play the major role in the drug binding. The binding average distance between furosemide and hCA II was estimated on the basis of the theory of Förster energy transfer. Decrease of protein surface hydrophobicity was also documented upon furosemide binding. Chemical modification of hCA II using N-bromosuccinimide indicated decrease of the number of accessible tryptophans in the presence of furosemide. CD results suggested the occurance of some alterations in α-helical content as well as tertiary structure of hCA II upon drug binding.
authors.affiliation: Department of Biology, Faculty of Science, Razi University, Kermanshah, Iran.
authors.forename: Samira
authors.initials: S
authors.lastname: Ranjbar
authors.forename: Sirous
authors.initials: S
authors.lastname: Ghobadi
authors.forename: Reza
authors.initials: R
authors.lastname: Khodarahmi
authors.forename: Houshang
authors.initials: H
authors.lastname: Nemati
mesh_terms: D001665:Binding Sites; D024402:Carbonic Anhydrase II / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004735:Energy Transfer; D005665:Furosemide / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D011485:Protein Binding; D011487:Protein Conformation / Q000187:drug effects; D013057:Spectrum Analysis; D014364:Tryptophan
chemical_list: D005665:Furosemide; D014364:Tryptophan; D024402:Carbonic Anhydrase II