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
1AKD 1997-05-16T00:00:00+0000 1.8 CYTOCHROME P450CAM FROM PSEUDOMONAS PUTIDA, COMPLEXED WITH 1S-CAMPHOR
1C8J 2000-05-31T00:00:00+0000 2.1 CRYSTAL STRUCTURE OF CYTOCHROME P450CAM MUTANT (F87W/Y96F)
1CP4 1991-06-04T00:00:00+0000 1.9 FORMATION, CRYSTAL STRUCTURE, AND REARRANGEMENT OF A CYTOCHROME P450-CAM IRON-PHENYL COMPLEX
1DZ4 2000-02-16T00:00:00+0000 1.6 ferric p450cam from pseudomonas putida
1DZ6 2000-02-18T00:00:00+0000 1.9 ferrous p450cam from pseudomonas putida
1DZ8 2000-02-18T00:00:00+0000 1.9 oxygen complex of p450cam from pseudomonas putida
1DZ9 2000-02-18T00:00:00+0000 1.9 Putative oxo complex of P450cam from Pseudomonas putida
1GEB 2000-11-01T00:00:00+0000 2.03 X-RAY CRYSTAL STRUCTURE AND CATALYTIC PROPERTIES OF THR252ILE MUTANT OF CYTOCHROME P450CAM
1GEK 2000-11-13T00:00:00+0000 1.7 STRUCTURAL CHARACTERIZATION OF N-BUTYL-ISOCYANIDE COMPLEXES OF CYTOCHROMES P450NOR AND P450CAM
1GEM 2000-11-13T00:00:00+0000 2.0 STRUCTURAL CHARACTERIZATION OF N-BUTYL-ISOCYANIDE COMPLEXES OF CYTOCHROMES P450NOR AND P450CAM

Relevant UniProtKB Entries

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