Differential Contribution of Active Site Residues in Substrate Recognition Sites 1 and 5 to Cytochrome P450 2C8 Substrate Selectivity and Regioselectivity†

Biochemistry ◽  
2004 ◽  
Vol 43 (24) ◽  
pp. 7834-7842 ◽  
Author(s):  
Oranun Kerdpin ◽  
David J. Elliot ◽  
Sanford L. Boye ◽  
Donald J. Birkett ◽  
Krongtong Yoovathaworn ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Active Site ◽  
Substrate Recognition ◽  
Substrate Selectivity ◽  
Active Site Residues ◽  
Recognition Sites ◽  
Cytochrome P450 2C8 ◽  
Differential Contribution

The synthesis of a new active-site analogue of cytochrome P450 carrying substrate recognition sites and athiolate ligand

1996 ◽  
Vol 113 (1-2) ◽  
pp. 393-402 ◽  
Author(s):  
Hamed Aissaoui ◽  
Sandro Ghirlanda ◽  
Christoph Gmür ◽  
Wolf-Dietrich Woggon
Keyword(s):  
Cytochrome P450 ◽  
Active Site ◽  
Substrate Recognition ◽  
Recognition Sites

scholarly journals Investigation of the role of cytochrome P450 2B4 active site residues in substrate metabolism based on crystal structures of the ligand-bound enzyme

2006 ◽  
Vol 455 (1) ◽  
pp. 61-67 ◽  
Author(s):  
Cynthia E. Hernandez ◽  
Santosh Kumar ◽  
Hong Liu ◽  
James R. Halpert
Keyword(s):  
Cytochrome P450 ◽  
Crystal Structures ◽  
Active Site ◽  
Active Site Residues ◽  
Substrate Metabolism ◽  
Cytochrome P450 2B4

scholarly journals Specificity and mechanism of carbohydrate demethylation by cytochrome P450 monooxygenases

2018 ◽  
Vol 475 (23) ◽  
pp. 3875-3886 ◽  
Author(s):  
Craig S. Robb ◽  
Lukas Reisky ◽  
Uwe T. Bornscheuer ◽  
Jan-Hendrik Hehemann
Keyword(s):  
Cytochrome P450 ◽  
Active Site ◽  
Substrate Recognition ◽  
High Energy ◽  
Cytochrome P450 Monooxygenases ◽  
P450 Monooxygenases ◽  
High Energy Barrier ◽  
Molecular Determinants ◽  
Cytochrome P450 Family ◽  
Carbohydrate Ligand

Degradation of carbohydrates by bacteria represents a key step in energy metabolism that can be inhibited by methylated sugars. Removal of methyl groups, which is critical for further processing, poses a biocatalytic challenge because enzymes need to overcome a high energy barrier. Our structural and computational analysis revealed how a member of the cytochrome P450 family evolved to oxidize a carbohydrate ligand. Using structural biology, we ascertained the molecular determinants of substrate specificity and revealed a highly specialized active site complementary to the substrate chemistry. Invariance of the residues involved in substrate recognition across the subfamily suggests that they are critical for enzyme function and when mutated, the enzyme lost substrate recognition. The structure of a carbohydrate-active P450 adds mechanistic insight into monooxygenase action on a methylated monosaccharide and reveals the broad conservation of the active site machinery across the subfamily.

Topological role of cytochrome P450 2D6 active site residues

2006 ◽  
Vol 447 (1) ◽  
pp. 53-58 ◽  
Author(s):  
Robert A.B. van Waterschoot ◽  
Peter H.J. Keizers ◽  
Chris de Graaf ◽  
Nico P.E. Vermeulen ◽  
Richard A. Tschirret-Guth
Keyword(s):  
Cytochrome P450 ◽  
Active Site ◽  
Cytochrome P450 2D6 ◽  
Active Site Residues ◽  
P450 2D6

scholarly journals CYP2E1 active site residues in substrate recognition sequence 5 identified by photoaffinity labeling and homology modeling

2007 ◽  
Vol 459 (1) ◽  
pp. 59-69 ◽  
Author(s):  
Samuel L. Collom ◽  
Arvind P. Jamakhandi ◽  
Alan J. Tackett ◽  
Anna Radominska-Pandya ◽  
Grover P. Miller
Keyword(s):  
Homology Modeling ◽  
Active Site ◽  
Substrate Recognition ◽  
Photoaffinity Labeling ◽  
Active Site Residues ◽  
Recognition Sequence

Reassessment of Cytochrome P450 2B2:  Catalytic Specificity and Identification of Four Active Site Residues†

Biochemistry ◽  
1997 ◽  
Vol 36 (39) ◽  
pp. 11697-11706 ◽  
Author(s):  
Sharon M. Strobel ◽  
James R. Halpert
Keyword(s):  
Cytochrome P450 ◽  
Active Site ◽  
Active Site Residues

scholarly journals Phe120 contributes to the regiospecificity of cytochrome P450 2D6: mutation leads to the formation of a novel dextromethorphan metabolite

2004 ◽  
Vol 380 (2) ◽  
pp. 353-360 ◽  
Author(s):  
Jack U. FLANAGAN ◽  
Jean-Didier MARÉCHAL ◽  
Richard WARD ◽  
Carol A. KEMP ◽  
Lesley A. McLAUGHLIN ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Active Site ◽  
Structural Model ◽  
Side Chain ◽  
Cytochrome P450 2D6 ◽  
Active Site Residues ◽  
Bacterial Membranes ◽  
Human Cytochrome ◽  
Methionine Residues ◽  
P450 2D6

Although the residues that determine the preference of CYP2D6 (cytochrome P450 2D6) for compounds containing a basic nitrogen are well characterized, the contribution of other active site residues to substrate binding and orientation is less well understood. Our structural model of CYP2D6 identifies the aromatic residue Phe120 as a likely major feature of the active site. To examine the role of Phe120, mutants of CYP2D6 in which this residue has been substituted by alanine, leucine, tyrosine, serine, histidine, tryptophan or methionine residues have been prepared in bacterial membranes co-expressing human cytochrome NADPH cytochrome P450 oxidoreductase. The mutants have been characterized using the prototypical bufuralol 1´ hydroxylase and dextromethorphan O- and N-demethylase activities of CYP2D6. Larger effects on Km values are observed for dextromethorphan O-demethylation than for bufuralol 1´ hydroxylation, indicating that the Phe120 side chain is more important in dextromethorphan than in bufuralol binding. A role for this side chain in determining the regiospecificity of substrate oxidation was indicated by changes in the relative rates of O- and N-demethylation of dextromethorphan and, notably, by the formation of 7-hydroxy dextromethrophan, a novel dextromethorphan metabolite, in mutants in which it had been substituted. Computational studies of dextromethorphan binding to the active site of the Phe120→Ala mutant were carried out to throw light on the way in which the removal of this side chain leads to different modes of ligand binding.

Cytochrome P450: Probes of Active Site Residues

1993 ◽  
pp. 183-194
Author(s):  
L. S. Kaminsky ◽  
R. S. Obach ◽  
M. J. Fasco
Keyword(s):  
Cytochrome P450 ◽  
Active Site ◽  
Active Site Residues
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