ライフサイエンスコーパス: CYP101

1 CYP101 (cytochrome P450cam) catalyses the oxidation of c

2 CYP101 forms a specific electron transfer complex with i

3 H, 15N correlations in perdeuterated [U-15N] CYP101 were monitored as a function of K+ concentration

4 Chemical shifts of perdeuterated [U-15N]CYP101 backbone amide (NH) resonances were monitored as

5 sites on Pdx for putidaredoxin reductase and CYP101.

6 tron of the catalytic cycle to camphor-bound CYP101[FeO2](2+) Judging by the appearance of an absorba

7 ication of the residues strongly affected by CYP101 binding.

8 te characterization of the complex formed by CYP101 and Pdx.

9 been shown to be an effector for turnover by CYP101.

10 he mapping of two bacterial P450s, P450 cam (CYP101) and P450 BM-3 (CYP102), identified the substrate

11 bon hydroxylation by cytochromes P-450(cam) (CYP101) and P-450(BM3) (CYP102).

12 he camphor hydroxylase cytochrome P450(cam) (CYP101) catalyzes the 5-exo hydroxylation of camphor in

13 he camphor hydroxylase cytochrome P450(cam) (CYP101) depend upon the oxidation and ligation state of

14 he camphor hydroxylase cytochrome P450(cam) (CYP101) obtained from residual dipolar coupling (RDC)-re

15 iron ligand, the C357H mutant of P450(cam) (CYP101) was characterized by resonance Raman, UV, circul

16 ing the oxidation of norcarane by P450(cam) (CYP101), P450(BM3) (CYP102), CYP2B1, and CYP2E1.

17 l electron transfer to cytochrome P450(cam) (CYP101).

18 -Pro89 peptide bond in cytochrome P450(cam) (CYP101).

19 is supplied directly to purified cytochrome CYP101 (P450cam; EC 1.14.15.1) through its natural redox

20 of substrate- and carbonmonoxy-bound ferrous CYP101 with sequence-specific Pdx-induced structural per

21 the physiological reductant and effector for CYP101 in the enzymatic reaction involving conversion of

22 H, (15)N, and (13)C resonance assignments in CYP101 that permit a more complete characterization of t

23 NMR-detected perturbations in CYP101 upon Pdx binding encompass regions of the CYP101

24 Cryoradiolysis of the mutant CYP101 oxyferrous complexes further indicates a specific

25 hexachloroethane dehalogenation activity of CYP101 has been investigated by mutagenesis.

26 lt bridge between Asp38 of Pdx and Arg112 of CYP101, while at the same time identifying key features

27 or is to populate the active conformation of CYP101 to prevent uncoupling.

28 ydroxycamphor) by enforcing conformations of CYP101 that prevent loss of substrate and/or intermediat

29 ytochrome b5, a nonphysiological effector of CYP101, were investigated by NMR spectroscopy and compar

30 the Pdx binding site on the proximal face of CYP101 and the site of isomerization is described.

31 ading to the catalytically competent form of CYP101 upon binding of the effector protein putidaredoxi

32 ed at the 1H resonance of the 8-CH3 group of CYP101-bound camphor upon addition of cytochrome b5, a p

33 located across from the heme-binding loop of CYP101 and forms non-polar contacts with several residue

34 pin-state equilibrium in the L358P mutant of CYP101 is more sensitive to K+ concentration than WT CYP

35 was 36 nmol of 5-exo-hydroxycamphor/nmol of CYP101 per min.

36 n that acts as the effector and reductant of CYP101 in vivo.

37 l spectra of the reduced oxygenated state of CYP101 and show that the primary intermediate, a hydrope

38 esidues in the vicinity of the heme group on CYP101, pointing to a potentially important role in comp

39 ed to CYP101[FeO(2+)por(*+)] (compound 1) or CYP101[FeO(2+)] (compound 2).

40 nzenes by mutants of the haem mono-oxygenase CYP101 (cytochrome P450(cam)) from Pseudomonas putida wi

41 lytic one-electron reduction of the oxy-P450 CYP101 complex.

42 tase), and that of a bacterial soluble P450, CYP101 when bound with their most common substrates, and

43 geted dehydration of the cytochrome P450cam (CYP101) distal pocket through mutagenesis of a distal po

44 Cytochrome P450cam (CYP101) from Pseudomonas putida is unusual among P450 en

45 putidaredoxin (Pdx), and cytochrome P450cam (CYP101) from the bacterium Pseudomonas putida has been i

46 uctural perturbations in cytochrome P450cam (CYP101) induced by the soluble fragment of cytochrome b5

47 Cytochrome P450cam (CYP101) is a prokaryotic monooxygenase that requires two

48 The camphor monoxygenase cytochrome P450cam (CYP101) requires potassium ion (K+) to drive formation o

49 ts redox partners in the cytochrome P450cam (CYP101) system was investigated by site-directed mutagen

50 exes designed to bind to cytochrome p450cam (CYP101).

51 -free) and camphor-bound cytochrome P450cam (CYP101).

52 in the binding of Pdx to cytochrome P450cam (CYP101).

53 experimental structure for the oxidized Pdx-CYP101 complex, a combined approach using orientational

54 The new NMR structure of the Pdx-CYP101 complex agrees well with results from previous mu

55 ed (CYP-S) and camphor- and CO-bound reduced CYP101 (CYP-S-CO).

56 ssociation of cytochrome b5 with the reduced CYP101-camphor-carbon monoxide complex (CYP-S-CO) pertur

57 sorbance maximum at 440 nm, we conclude that CYP101[FeOOH](2+) (compound 0) accumulates within 5 mus

58 nces that Pdx does, including regions of the CYP101 molecule implicated in substrate access and orien

59 ite, including in particular a region of the CYP101 molecule that has been implicated in substrate ac

60 01 upon Pdx binding encompass regions of the CYP101 remote from the putative Pdx binding site, includ

61 on also associated with the formation of the CYP101 x Pdx complex, albeit with larger perturbations.

62 The results suggested that the CYP101 active site shows good match with hexachloroethan

63 ly for Pdx in terms of binding affinities to CYP101, NMR spectral differences, and dynamic properties

64 nsient absorbance bands could be assigned to CYP101[FeO(2+)por(*+)] (compound 1) or CYP101[FeO(2+)] (

65 l residue of Pdx and critical for binding to CYP101, is located across from the heme-binding loop of

66 the subunit binding and electron transfer to CYP101.

67 Wild-type CYP101 had low activity for the oxidation of dichloroben

68 dation systems could be constructed in which CYP101 mutants convert the inert polychlorinated benzene

69 metal cluster region of Pdx in complex with CYP101 have also been mapped for the first time using (1

70 conformations enforced by Pdx binding in WT CYP101.

71 s more sensitive to K+ concentration than WT CYP101, consistent with a hypothesis that L358P preferen