The regulation of cytochrome P450 activity is often achieved by structural transitions induced by substrate binding. We describe the conformational transition experienced upon binding by the P450 OleP, an epoxygenase involved in oleandomycin biosynthesis. OleP bound to the substrate analog 6DEB crystallized in 2 forms: one with an ensemble of open and closed conformations in the asymmetric unit and another with only the closed conformation. Characterization of OleP-6DEB binding kinetics, also using the P450 inhibitor clotrimazole, unveiled a complex binding mechanism that involves slow conformational rearrangement with the accumulation of a spectroscopically detectable intermediate where 6DEB is bound to open OleP. Data reported herein provide structural snapshots of key precatalytic steps in the OleP reaction and explain how structural rearrangements induced by substrate binding regulate activity.

Substrate-induced conformational change in cytochrome P450 OleP / Parisii, Giacomo; Montemiglio, LINDA CELESTE; Giuffrè, Alessandro; Macone, Alberto; Scaglione, Antonella; Cerutti, Gabriele; Exertier, Cecile; Savino, Carmelinda; Vallone, Beatrice. - In: THE FASEB JOURNAL. - ISSN 0892-6638. - 33:2(2019), pp. 1787-1800. [10.1096/fj.201800450RR]

Substrate-induced conformational change in cytochrome P450 OleP

Montemiglio Linda Celeste
Secondo
Investigation
;
Giuffrè Alessandro
Membro del Collaboration Group
;
Macone Alberto
Investigation
;
Scaglione Antonella
Membro del Collaboration Group
;
Cerutti Gabriele
Investigation
;
EXERTIER, CECILE
Investigation
;
Savino Carmelinda
Penultimo
Membro del Collaboration Group
;
Vallone Beatrice
Ultimo
Supervision
2019

Abstract

The regulation of cytochrome P450 activity is often achieved by structural transitions induced by substrate binding. We describe the conformational transition experienced upon binding by the P450 OleP, an epoxygenase involved in oleandomycin biosynthesis. OleP bound to the substrate analog 6DEB crystallized in 2 forms: one with an ensemble of open and closed conformations in the asymmetric unit and another with only the closed conformation. Characterization of OleP-6DEB binding kinetics, also using the P450 inhibitor clotrimazole, unveiled a complex binding mechanism that involves slow conformational rearrangement with the accumulation of a spectroscopically detectable intermediate where 6DEB is bound to open OleP. Data reported herein provide structural snapshots of key precatalytic steps in the OleP reaction and explain how structural rearrangements induced by substrate binding regulate activity.
2019
6DEB; X-ray crystallography; multistep binding kinetics; spectroscopic intermediate; structural transition
01 Pubblicazione su rivista::01a Articolo in rivista
Substrate-induced conformational change in cytochrome P450 OleP / Parisii, Giacomo; Montemiglio, LINDA CELESTE; Giuffrè, Alessandro; Macone, Alberto; Scaglione, Antonella; Cerutti, Gabriele; Exertier, Cecile; Savino, Carmelinda; Vallone, Beatrice. - In: THE FASEB JOURNAL. - ISSN 0892-6638. - 33:2(2019), pp. 1787-1800. [10.1096/fj.201800450RR]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1175862
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