Thermodynamic and structural stability of cytochrome c oxidase from Paracoccus denitrificans.


The structural stability of the integral membrane protein cytochrome c oxidase from Paracoccus denitrificans has been measured by high-sensitivity differential scanning calorimetry and Fourier transform infrared spectroscopy. Contrary to the mammalian enzyme or the yeast enzyme, which are composed of 13 subunits, the bacterial enzyme has only three or four subunits, thus providing a unique opportunity to examine the magnitude of the forces that stabilize this enzyme and to establish accurate structural assignments of events observed calorimetrically. In this paper, experiments have been performed with the wild-type enzyme and with a mutant enzyme lacking subunit III. Our results show that subunits I and II form a highly cooperative complex which denatures as a single cooperative unit at 67 degrees C, while subunit III is less stable and denatures 20 degrees C earlier. Reduction of the enzyme causes a large increase in the stability of subunits I and II but has absolutely no effect on subunit III. Despite the lack of a strong interaction between subunit III and the catalytic subunits, the absence of subunit III leads to a turnover-induced loss of electron-transfer activity. The magnitude of the energetic parameters and the infrared spectroscopic experiments indicate that the enzyme does not completely unfold upon thermal denaturation and that significant degrees of structure are preserved. The amount of native alpha-helix structure, which is 45% in the native state, decreases only to 30% after thermal denaturation. Presumably, the residual helical structure existing after thermal denaturation belongs to the intramembranous portions of the protein. The calorimetric behavior of subunit III does not fully conform to that expected for a highly alpha-helical membrane protein. The picture that emerges from these experiments is that, in the temperature-denatured form of the enzyme, most of the extramembranous structural elements are denatured while most of the intramembranous secondary structure is maintained even though native tertiary interactions appear to be disrupted. Study holds ProTherm entries: 4515, 4516 Extra Details: the transition is from native to intermediate integral membrane protein; structural assignments;,cooperative complex; catalytic subunits; alpha-helical

Submission Details

ID: Af6Pxg334

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:25 p.m.

Version: 1

Publication Details
Haltia T;Semo N;Arrondo JL;Goñi FM;Freire E,Biochemistry (1994) Thermodynamic and structural stability of cytochrome c oxidase from Paracoccus denitrificans. PMID:8068652
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 UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Cytochrome c oxidase subunit 1-beta P98002 COX1B_PARDE
90.5 Cytochrome c oxidase subunit 1-beta P08305 COX1A_PARDE
100.0 Cytochrome c oxidase subunit 1-beta Q00502 COX1_PARVE
92.9 C Cytochrome c oxidase subunit 1-beta P18525 HVM54_MOUSE
100.0 B Cytochrome c oxidase subunit 1-beta P08306 COX2_PARDE