Functional and structural changes of human erythrocyte catalase induced by cimetidine: proposed model of binding.
In erythrocyte, catalase plays an important role to protect cells from hydrogen peroxide toxicity. Hydrogen peroxide is a byproduct compound which is produced during metabolic pathway of cells. Cimetidine, a histamine H2 receptor antagonist, is used for gastrointestinal tract diseases and prevents the extra release of gastric acid. In this study, the effect of cimetidine on the activity of human erythrocyte catalase was investigated. Erythrocytes were broken by hypotonic solution. The supernatant was used for catalase assay and kinetics study. Lineweaver-Burk plot was performed to determine the type of inhibition. The kinetics data revealed that cimetidine inhibited the catalase activity by mixed inhibition. The IC50 (1.54 μM) and Ki (0.45 μM) values of cimetidine determined that the drug was bound to the enzyme with high affinity. Circular dichroism and fluorescence measurement showed that the binding of cimetidine to the enzyme affected the content of secondary structure of the enzyme as well as its conformational changes. Docking studies were carried out to detect the site in which the drug was bound to the enzyme. Molecular modeling and energy calculation of the binding showed that the cyanoguanidine group of the drug connected to Asp59 via two hydrogen bonds, while the imidazole group of the drug interacted with Phe64 in the enzyme by a hydrophobic interaction. In conclusion, cimetidine could bind to human erythrocyte catalase, and its interaction caused functional and conformational changes in the enzyme.
Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran.
Fatemeh
F
Yazdi
Dariush
D
Minai-Tehrani
Mahboubeh
M
Jahngirvand
Ali
A
Almasirad
Zahra
Z
Mousavi
Masoudeh
M
Masoud
Hamidreza
H
Mollasalehi
D001665:Binding Sites; D002374:Catalase / Q000737:chemistry / Q000378:metabolism*; D002927:Cimetidine / Q000494:pharmacology*; D004912:Erythrocytes / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D062105:Molecular Docking Simulation; D011485:Protein Binding / Q000187:drug effects; D011487:Protein Conformation / Q000187:drug effects; D013329:Structure-Activity Relationship
D002927:Cimetidine; D006861:Hydrogen Peroxide; D002374:Catalase