Apocytochrome P450cam is a native protein with some intermediate-like properties.


Abstract

Holo- and apocytochrome P450cam were studied by differential scanning calorimetry (DSC), limited proteolysis, second-derivative spectroscopy, circular dichroism, and size-exclusion chromatography. The holoprotein shows three folding units (domains) in DSC. The prosthetic group is related to the most unstable domain, which has a thermal transition at 41.9 degrees C. Compared with the holoprotein, apocytochrome P450cam has a reduced helix content. The protein is compact as judged by the Stokes radius and is still able to undergo a two-state transition. However, the enthalpy change at thermal melting is reduced from 980 kJ/mol for the holoprotein to 135 kJ/mol for the apo form. Parts of the molecule have a destabilized tertiary structure. This is indicated by second-derivative spectroscopy, circular dichroism in the near-ultraviolet region, and a high susceptibility to proteolytic digestion. Apocytochrome P450cam is considered a native protein with the extremely low stability of delta G = 7.5 kJ/mol, thus showing at the same time intermediate-like properties. The importance of the properties for in vivo folding are discussed. Study holds ProTherm entries: 4549 Extra Details: KCN(1 mM) was added in the experiment folding units; helix content; Stokes radius; susceptibility;,tertiary structure

Submission Details

ID: dQ4Z6T9m

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:26 p.m.

Version: 1

Publication Details
Pfeil W;Nölting BO;Jung C,Biochemistry (1993) Apocytochrome P450cam is a native protein with some intermediate-like properties. PMID:8364032
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
2LQD 2012-05-09 Reduced and CO-bound cytochrome P450cam (CYP101A1)
2L8M 2011-02-16 Reduced and CO-bound cytochrome P450cam (CYP101A1)
2M56 2013-08-21 The structure of the complex of cytochrome P450cam and its electron donor putidaredoxin determined by paramagnetic NMR spectroscopy
4L4E 2013-07-31 1.26 Structure of cyanide and camphor bound P450cam mutant L358A/K178G
4L4F 2013-07-31 1.29 Structure of cyanide and camphor bound P450cam mutant L358A/K178G/D182N
2ZWU 2009-02-17 1.3 Crystal Structure of Camphor Soaked Ferric Cytochrome P450cam
2ZWT 2009-02-17 1.35 Crystal Structure of Ferric Cytochrome P450cam
1YRC 2005-02-15 1.4 X-ray Crystal Structure of hydrogenated Cytochrome P450cam
1RE9 2004-11-16 1.45 CRYSTAL STRUCTURE OF CYTOCHROME P450-CAM WITH A FLUORESCENT PROBE D-8-AD (ADAMANTANE-1-CARBOXYLIC ACID-5-DIMETHYLAMINO-NAPHTHALENE-1-SULFONYLAMINO-OCTYL-AMIDE)
2FE6 2006-03-14 1.5 P450CAM from Pseudomonas putida reconstituted with manganic protoporphyrin IX
3L61 2010-04-21 1.5 Crystal structure of substrate-free P450cam at 200 mM [K+]
3L63 2010-04-21 1.5 Crystal structure of camphor-bound P450cam at low [K+]
2H7Q 2006-10-31 1.5 Cytochrome P450cam complexed with imidazole
2ZUI 2009-10-20 1.5 Crystal Structure of Camphor-soaked Ferric Cytochrome P450cam Mutant (D297N)
5IK1 2016-05-04 1.53 Open state of P450cam after soaking in camphor
3FWG 2009-03-03 1.55 Ferric camphor bound Cytochrome P450cam, Arg365Leu, Glu366Gln, monoclinic crystal form
2ZAW 2008-01-01 1.55 Crystal Structure of Ferric Cytochrome P450cam Reconstituted with 6-Methyl-6-depropionated Hemin
2ZUH 2009-10-20 1.55 Crystal Structure of Camphor-soaked Ferric Cytochrome P450cam Mutant (D297A)
2A1O 2005-07-05 1.55 Crystal structure of ferrous dioxygen complex of T252A cytochrome P450cam
4L4G 2013-07-31 1.55 Structure of cyanide and camphor bound P450cam mutant L358P/K178G
1QMQ 2000-10-06 1.55 Optical detection of cytochrome P450 by sensitizer-linked substrates
2ZUJ 2009-10-20 1.6 Crystal Structure of Camphor-soaked Ferric Cytochrome P450cam Mutant(D297L)
1PHB 1993-10-31 1.6 INHIBITOR-INDUCED CONFORMATIONAL CHANGE IN CYTOCHROME P450-CAM
2ZAX 2008-01-01 1.6 Crystal Structure of Ferric Cytochrome P450cam
1DZ4 2000-07-20 1.6 ferric p450cam from pseudomonas putida
1PHD 1993-10-31 1.6 CRYSTAL STRUCTURES OF METYRAPONE-AND PHENYLIMIDAZOLE-INHIBITED COMPLEXES OF CYTOCHROME P450-CAM
1PHF 1993-10-31 1.6 CRYSTAL STRUCTURES OF METYRAPONE-AND PHENYLIMIDAZOLE-INHIBITED COMPLEXES OF CYTOCHROME P450-CAM
1PHG 1993-10-31 1.6 CRYSTAL STRUCTURES OF METYRAPONE-AND PHENYLIMIDAZOLE-INHIBITED COMPLEXES OF CYTOCHROME P450-CAM
1PHE 1993-10-31 1.6 CRYSTAL STRUCTURES OF METYRAPONE-AND PHENYLIMIDAZOLE-INHIBITED COMPLEXES OF CYTOCHROME P450-CAM
1PHC 1993-10-31 1.6 CRYSTAL STRUCTURE OF SUBSTRATE-FREE PSEUDOMONAS PUTIDA CYTOCHROME P450
3WRH 2015-03-18 1.62 Crystal structure of P450cam
1PHA 1993-10-31 1.63 INHIBITOR-INDUCED CONFORMATIONAL CHANGE IN CYTOCHROME P450-CAM
2CPP 1987-07-16 1.63 HIGH-RESOLUTION CRYSTAL STRUCTURE OF CYTOCHROME P450-CAM
1K2O 2001-10-17 1.65 Cytochrome P450Cam with Bound BIS(2,2'-BIPYRIDINE)-(5-METHYL-2-2'-BIPYRIDINE)-C2-ADAMANTANE RUTHENIUM (II)
3P6X 2010-11-17 1.65 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog AdaC3-C8-Dans
3OIA 2010-11-17 1.65 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog AdaC1-C8GluEtg-Bio
3WRL 2015-03-18 1.65 Crystal structure of P450cam
1YRD 2005-02-15 1.7 X-ray crystal structure of PERDEUTERATED Cytochrome P450cam
5GXG 2017-01-18 1.7 High-resolution crystal structure of the electron transfer complex of cytochrome p450cam with putidaredoxin
2FER 2006-03-14 1.7 P450CAM from Pseudomonas putida reconstituted with manganic protoporphyrin IX
3P6N 2010-11-17 1.7 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog AdaC1-C8-Dans
2FEU 2006-03-14 1.7 P450CAM from Pseudomonas putida reconstituted with manganic protoporphyrin IX
3P6U 2010-11-17 1.7 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog AdaC3-C6-Dans
1GEK 2000-12-06 1.7 STRUCTURAL CHARACTERIZATION OF N-BUTYL-ISOCYANIDE COMPLEXES OF CYTOCHROMES P450NOR AND P450CAM
3L62 2010-04-21 1.7 Crystal structure of substrate-free P450cam at low [K+]
3OL5 2010-11-17 1.75 Crystal Structure of Cytochrome P450cam crystallized with a tethered substrate analog 3OH-AdaC1-C8-Dans
5CP4 1998-09-16 1.75 CRYOGENIC STRUCTURE OF P450CAM
2QBN 2007-12-25 1.75 Crystal structure of ferric G248V cytochrome P450cam
1T87 2004-05-25 1.8 Crystal Structure of the Ferrous CO-bound Cytochrome P450cam (C334A)
1AKD 1997-11-19 1.8 CYTOCHROME P450CAM FROM PSEUDOMONAS PUTIDA, COMPLEXED WITH 1S-CAMPHOR
2QBM 2007-12-25 1.8 Crystal structure of the P450cam G248T mutant in the cyanide bound state
1RF9 2004-11-16 1.8 Crystal structure of cytochrome P450-cam with a fluorescent probe D-4-AD (Adamantane-1-carboxylic acid-5-dimethylamino-naphthalene-1-sulfonylamino-butyl-amide)
2QBL 2007-12-25 1.8 Crystal structure of ferric G248T cytochrome P450cam
1T85 2004-06-01 1.8 Crystal Structure of the Ferrous CO-bound Cytochrome P450cam Mutant (L358P/C334A)
1O76 2002-12-19 1.8 CYANIDE COMPLEX OF P450CAM FROM PSEUDOMONAS PUTIDA
2Z97 2008-09-23 1.8 Crystal Structure of Ferric Cytochrome P450cam Reconstituted with 7-Methyl-7-depropionated Hemin
3FWF 2009-03-03 1.83 Ferric camphor bound cytochrome P450cam containing a Selenocysteine as the 5th heme ligand, monoclinic crystal form
3WRJ 2015-03-18 1.85 Crystal structure of P450cam
3P6P 2010-11-17 1.9 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog AdaC1-C6-Bio
6CP4 1998-09-16 1.9 P450CAM D251N MUTANT
1T88 2004-05-25 1.9 Crystal Structure of the Ferrous Cytochrome P450cam (C334A)
3CPP 1990-04-15 1.9 CRYSTAL STRUCTURE OF THE CARBON MONOXY-SUBSTRATE-CYTOCHROME P450-CAM TERNARY COMPLEX
1T86 2004-05-25 1.9 Crystal Structure of the Ferrous Cytochrome P450cam Mutant (L358P/C334A)
3P6T 2010-11-17 1.9 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog AdaC2-C8-Dans
3FWJ 2009-03-03 1.9 Ferric camphor bound Cytochrome P450cam containing a selenocysteine as the 5th heme ligand, orthorombic crystal form
6CPP 1991-07-15 1.9 CRYSTAL STRUCTURES OF CYTOCHROME P450-CAM COMPLEXED WITH CAMPHANE, THIOCAMPHOR, AND ADAMANTANE: FACTORS CONTROLLING P450 SUBSTRATE HYDROXYLATION
1DZ6 2000-03-30 1.9 ferrous p450cam from pseudomonas putida
1DZ8 2000-03-31 1.9 oxygen complex of p450cam from pseudomonas putida
2A1N 2005-07-05 1.9 Crystal structure of ferrous dioxygen complex of D251N cytochrome P450cam
2QBO 2007-12-25 1.9 Crystal structure of the P450cam G248V mutant in the cyanide bound state
1CP4 1993-01-15 1.9 FORMATION, CRYSTAL STRUCTURE, AND REARRANGEMENT OF A CYTOCHROME P450-CAM IRON-PHENYL COMPLEX
1DZ9 2000-03-30 1.9 Putative oxo complex of P450cam from Pseudomonas putida
3P6Q 2010-11-17 1.95 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog AdaC2-Etg-Boc
3WRM 2015-03-18 1.95 Crystal structure of P450cam
4EK1 2012-07-25 1.97 Crystal Structure of Electron-Spin Labeled Cytochrome P450cam
5WK7 2017-09-06 1.98 P450cam mutant R186A
5WK9 2017-09-06 1.98 R186AP450cam with CN and camphor
1GEM 2000-12-06 2.0 STRUCTURAL CHARACTERIZATION OF N-BUTYL-ISOCYANIDE COMPLEXES OF CYTOCHROMES P450NOR AND P450CAM
1IWJ 2002-06-05 2.0 Putidaredoxin-Binding Stablilizes an Active Conformer of Cytochrome P450cam in its Reduced State; Crystal Structure of Mutant(109K) Cytochrome P450cam
3P6V 2010-11-17 2.0 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog 3Et-AdaC1-Etg-Boc
1IWK 2002-06-05 2.0 Putidaredoxin-Binding Stablilizes an Active Conformer of Cytochrome P450cam in its Reduced State; Crystal Structure of Mutant(112K) Cytochrome P450cam
3P6O 2010-11-17 2.0 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog AdaC1-Etg-Dans
3P6M 2010-11-17 2.0 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog AdaC1-C8-Dans
1IWI 2002-06-05 2.0 Putidaredoxin-Binding Stablilizes an Active Conformer of Cytochrome P450cam in its Reduced State; Crystal Structure of Cytochrome P450cam
4KKY 2014-05-21 2.0 Crystal structure of N-(1-Pyrene)acetamide labeled P450cam in substrate bound form.
7CPP 1991-07-15 2.0 THE STRUCTURAL BASIS FOR SUBSTRATE-INDUCED CHANGES IN REDOX POTENTIAL AND SPIN EQUILIBRIUM IN CYTOCHROME P450(CAM)
1UYU 2005-03-07 2.0 Xenon COMPLEX OF wildtype P450CAM FROM PSEUDOMONAS PUTIDA
3P6S 2010-11-17 2.0 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog AdaC2-C8-Dans
1GEB 2000-11-15 2.03 X-RAY CRYSTAL STRUCTURE AND CATALYTIC PROPERTIES OF THR252ILE MUTANT OF CYTOCHROME P450CAM
5CPP 1991-07-15 2.08 THE STRUCTURAL BASIS FOR SUBSTRATE-INDUCED CHANGES IN REDOX POTENTIAL AND SPIN EQUILIBRIUM IN CYTOCHROME P-450(CAM)
4JX1 2013-06-19 2.09 Crystal structure of reduced Cytochrome P450cam-putidaredoxin complex bound to camphor and 5-exo-hydroxycamphor
2CP4 1993-01-15 2.1 CRYSTAL STRUCTURE OF THE CYTOCHROME P450-CAM ACTIVE SITE MUTANT THR252ALA
4L4B 2013-07-31 2.1 Structure of L358A/K178G/D182N mutant of P450cam bound to camphor
2H7R 2006-10-31 2.1 L244A mutant of Cytochrome P450cam complexed with imidazole
1C8J 2001-05-09 2.1 CRYSTAL STRUCTURE OF CYTOCHROME P450CAM MUTANT (F87W/Y96F)
3P6W 2010-11-17 2.1 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog AdaC3-Etg-Boc
4L4A 2013-07-31 2.1 Structure of L358A/K178G mutant of P450cam bound to camphor
2FRZ 2007-01-16 2.1 Crystal Structure of Cytochrome P450cam mutant (F87W/Y96F/V247L/C334A)
4CP4 1993-01-15 2.1 CRYSTAL STRUCTURE OF THE CYTOCHROME P450-CAM ACTIVE SITE MUTANT THR252ALA
2A1M 2005-07-05 2.1 Crystal structure of ferrous dioxygen complex of wild-type cytochrome P450cam
2GQX 2007-04-17 2.1 Crystal structure of cytochrome p450cam mutant (f87w/y96f/l244a/v247l/c334a) with pentachlorobenzene
8CPP 1991-07-15 2.1 CRYSTAL STRUCTURES OF CYTOCHROME P450-CAM COMPLEXED WITH CAMPHANE, THIOCAMPHOR, AND ADAMANTANE: FACTORS CONTROLLING P450 SUBSTRATE HYDROXYLATION
3P6R 2010-11-17 2.1 Crystal Structure of Cytochrome P450cam crystallized in the presence of a tethered substrate analog 3OH-AdaC1-Etg-Boc
4L4D 2013-07-31 2.1 Structure of cyanide and camphor bound P450cam mutant L358A
4CPP 1991-07-15 2.11 CRYSTAL STRUCTURES OF CYTOCHROME P450-CAM COMPLEXED WITH CAMPHANE, THIOCAMPHOR, AND ADAMANTANE: FACTORS CONTROLLING P450 SUBSTRATE HYDROXYLATION
4L49 2013-07-31 2.13 Structure of L358A mutant of P450cam bound to camphor
2H7S 2006-10-31 2.15 L244A mutant of Cytochrome P450cam
6NBL 2019-03-06 2.15 Cytochrome P450cam-putidaredoxin complex bound to camphor and cyanide
4JWS 2013-06-19 2.15 Crystal structure of Cytochrome P450cam-putidaredoxin complex
4JWU 2013-06-19 2.2 Crystal structure of Cytochrome P450cam-putidaredoxin complex
1NOO 1996-03-08 2.2 CYTOCHROME P450-CAM COMPLEXED WITH 5-EXO-HYDROXYCAMPHOR
4G3R 2012-08-01 2.2 Crystal Structure of Nitrosyl Cytochrome P450cam
1J51 2002-01-23 2.2 CRYSTAL STRUCTURE OF CYTOCHROME P450CAM MUTANT (F87W/Y96F/V247L/C334A) WITH 1,3,5-TRICHLOROBENZENE
1GJM 2001-07-31 2.2 Covalent attachment of an electroactive sulphydryl reagent in the active site of cytochrome P450cam
4L4C 2013-07-31 2.2 Structure of L358P/K178G mutant of P450cam bound to camphor
1LWL 2002-12-18 2.2 Crystal Structure of Cytochrome P450-cam with a Fluorescent Probe D-8-Ad (Adamantane-1-carboxylic acid-5-dimethylamino-naphthalene-1-sulfonylamino-octyl-amide)
3CP4 1993-01-15 2.3 CRYSTAL STRUCTURE OF THE CYTOCHROME P450-CAM ACTIVE SITE MUTANT THR252ALA
1P2Y 2003-10-28 2.3 CRYSTAL STRUCTURE OF CYTOCHROME P450CAM IN COMPLEX WITH (S)-(-)-NICOTINE
2GR6 2007-04-17 2.3 Crystal structure of cytochrome p450cam mutant (f87w/y96f/l244a/v247l/c334a)
1MPW 2002-10-09 2.34 Molecular Recognition in (+)-a-Pinene Oxidation by Cytochrome P450cam
3FWI 2009-03-03 2.4 Ferric camphor bound Cytochrome P450cam containing a selenocysteine as the 5th heme ligand, tetragonal crystal form
3W9C 2013-08-21 2.5 Crystal structure of the electron transfer complex of cytochrome p450cam with putidaredoxin
3WRK 2015-03-18 2.61 Crystal structure of P450cam
1P7R 2003-10-28 2.85 CRYSTAL STRUCTURE OF REDUCED, CO-EXPOSED COMPLEX OF CYTOCHROME P450CAM WITH (S)-(-)-NICOTINE
3WRI 2015-03-18 2.9 Crystal structure of P450cam

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Camphor 5-monooxygenase P00183 CPXA_PSEPU