Proton relay network in P450cam formed upon docking of putidaredoxin.
Proteins
; 88(4): 558-572, 2020 04.
Article
en En
| MEDLINE
| ID: mdl-31597203
ABSTRACT
Cytochromes P450 are versatile heme-based enzymes responsible for vital life processes. Of these, P450cam (substrate camphor) has been most studied. Despite this, precise mechanisms of the key OâO cleavage step remain partly elusive to date; effects observed in various enzyme mutants remain partly unexplained. We have carried out extended (to 1000 ns) MM-MD and follow-on quantum mechanics/molecular mechanics computations, both on the well-studied FeOO state and on Cpd(0) (compound 0). Our simulations include (all camphor-bound) (a) WT (wild type), FeOO state. (b) WT, Cpd(0). (c) Pdx (Putidaredoxin, redox partner of P450)-docked-WT, FeOO state. (d) Pdx-docked WT, Cpd(0). (e) Pdx-docked T252A mutant, Cpd(0). Among our key findings:
(a) Effect of Pdx docking appears to go far beyond that indicated in prior studies it leads to specific alterations in secondary structure that create the crucial proton relay network. (b) Specific proton relay networks we identify are FeOO(H)â¯T252â¯nH 2 Oâ¯D251 in WT; FeOO(H)â¯nH 2 Oâ¯D251 in T252A mutant; both occur with Pdx docking. (c) Direct interaction of D251 with -FeOOH is, respectively, rare/frequent in WT/T252A mutant. (d) In WT, T252 is in the proton relay network. (e) Positioning of camphor appears significant when camphor is part of H-bonding network, second protonation appears to be facilitated.Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Protones
/
Proteínas Bacterianas
/
Alcanfor
/
Alcanfor 5-Monooxigenasa
/
Ferredoxinas
/
Hemo
Tipo de estudio:
Prognostic_studies
Idioma:
En
Revista:
Proteins
Asunto de la revista:
BIOQUIMICA
Año:
2020
Tipo del documento:
Article