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Topological role of cytochrome P450 2D6 active site residues.
Arch Biochem Biophys. 2006 Mar 01; 447(1):53-8.AB

Abstract

Recent reports have identified Phe120, Asp301, Thr309, and Glu216 as important residues in cytochrome P450 2D6 (CYP2D6) substrate binding and catalysis. Complementary homology models have located these amino acids within the binding pocket of CYP2D6 and in the present study we have used aryldiazenes to test these models and gain further insight in the role these amino acids have in maintaining the integrity of the active site cavity. When Phe120 was replaced to alanine, there was a significant increase in probe migration to pyrrole nitrogens C and D, in agreement with homology models which have located the phenyl side-chain of Phe120 above these two pyrrole rings. No changes in topology were observed with the D301Q mutant, supporting claims that in this mutant the electrostatic interactions with the B/C-loop are largely maintained and the loop retains its native orientation. The T309V mutation resulted in significant topological alteration suggesting that, in addition to its potential role in dioxygen activation, Thr309 plays an important structural role within the active site crevice. Replacement of Ile106 with Glu, engineered to cause electrostatic repulsion with Glu216, had a profound topological effect in the higher region within the active site cavity and impaired the catalytic activity towards CYP2D6 probe substrates.

Authors+Show Affiliations

LACDR/Division of Molecular Toxicology, Department of Pharmacochemistry, Vrije Universiteit, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

16466686

Citation

van Waterschoot, Robert A B., et al. "Topological Role of Cytochrome P450 2D6 Active Site Residues." Archives of Biochemistry and Biophysics, vol. 447, no. 1, 2006, pp. 53-8.
van Waterschoot RA, Keizers PH, de Graaf C, et al. Topological role of cytochrome P450 2D6 active site residues. Arch Biochem Biophys. 2006;447(1):53-8.
van Waterschoot, R. A., Keizers, P. H., de Graaf, C., Vermeulen, N. P., & Tschirret-Guth, R. A. (2006). Topological role of cytochrome P450 2D6 active site residues. Archives of Biochemistry and Biophysics, 447(1), 53-8.
van Waterschoot RA, et al. Topological Role of Cytochrome P450 2D6 Active Site Residues. Arch Biochem Biophys. 2006 Mar 1;447(1):53-8. PubMed PMID: 16466686.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Topological role of cytochrome P450 2D6 active site residues. AU - van Waterschoot,Robert A B, AU - Keizers,Peter H J, AU - de Graaf,Chris, AU - Vermeulen,Nico P E, AU - Tschirret-Guth,Richard A, Y1 - 2006/01/23/ PY - 2005/12/14/received PY - 2006/01/04/revised PY - 2006/01/05/accepted PY - 2006/2/10/pubmed PY - 2006/4/28/medline PY - 2006/2/10/entrez SP - 53 EP - 8 JF - Archives of biochemistry and biophysics JO - Arch Biochem Biophys VL - 447 IS - 1 N2 - Recent reports have identified Phe120, Asp301, Thr309, and Glu216 as important residues in cytochrome P450 2D6 (CYP2D6) substrate binding and catalysis. Complementary homology models have located these amino acids within the binding pocket of CYP2D6 and in the present study we have used aryldiazenes to test these models and gain further insight in the role these amino acids have in maintaining the integrity of the active site cavity. When Phe120 was replaced to alanine, there was a significant increase in probe migration to pyrrole nitrogens C and D, in agreement with homology models which have located the phenyl side-chain of Phe120 above these two pyrrole rings. No changes in topology were observed with the D301Q mutant, supporting claims that in this mutant the electrostatic interactions with the B/C-loop are largely maintained and the loop retains its native orientation. The T309V mutation resulted in significant topological alteration suggesting that, in addition to its potential role in dioxygen activation, Thr309 plays an important structural role within the active site crevice. Replacement of Ile106 with Glu, engineered to cause electrostatic repulsion with Glu216, had a profound topological effect in the higher region within the active site cavity and impaired the catalytic activity towards CYP2D6 probe substrates. SN - 0003-9861 UR - https://www.unboundmedicine.com/medline/citation/16466686/Topological_role_of_cytochrome_P450_2D6_active_site_residues_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-9861(06)00006-3 DB - PRIME DP - Unbound Medicine ER -