title: Kinetic and structural analysis of the inhibition of adenosine deaminase by acetaminophen.
abstract: Kinetic and thermodynamic studies have been made on the effect of acetaminophen on the activity and structure of adenosine deaminase in 50 mM sodium phosphate buffer pH 7.5, at two temperatures of 27 and 37 degrees C using UV spectrophotometry, circular dichroism (CD) and fluorescence spectroscopy. Acetaminophen acts as a competitive inhibitor at 27 degrees C (Ki = 126 microM) and an uncompetitive inhibitor at 37 degrees C (Ki = 214 microM). Circular dichroism studies do not show any considerable effect on the secondary structure of adenosine deaminase by increasing the temperature from 27 to 37 degrees C. However, the secondary structure of the protein becomes more compact at 37 degrees C in the presence of acetaminophen. Fluorescence spectroscopy studies show considerable change in the tertiary structure of the protein by increasing the temperature from 27 to 37 degrees C. Also, the fluorescence spectrum of the protein incubated with different concentrations of acetaminophen show different inhibition behaviors by the effector at the two temperatures.
authors.affiliation: Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran.
authors.forename: G
authors.initials: G
authors.lastname: Ataie
authors.forename: S
authors.initials: S
authors.lastname: Safarian
authors.forename: A
authors.initials: A
authors.lastname: Divsalar
authors.forename: A A
authors.initials: AA
authors.lastname: Saboury
authors.forename: A A
authors.initials: AA
authors.lastname: Moosavi-Movahedi
authors.forename: B
authors.initials: B
authors.lastname: Ranjbar
authors.forename: G
authors.initials: G
authors.lastname: Cristalli
authors.forename: S
authors.initials: S
authors.lastname: Mardanian
mesh_terms: D000082:Acetaminophen / Q000737:chemistry / Q000494:pharmacology*; D000243:Adenosine Deaminase / Q000378:metabolism; D058892:Adenosine Deaminase Inhibitors; D001667:Binding, Competitive; D002942:Circular Dichroism; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D007700:Kinetics; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship; D013696:Temperature; D013816:Thermodynamics
chemical_list: D058892:Adenosine Deaminase Inhibitors; D004791:Enzyme Inhibitors; D000082:Acetaminophen; D000243:Adenosine Deaminase