Differential scanning calorimetry of bovine rhodopsin in rod-outer-segment disk membranes.


Abstract

Rhodopsin-containing retinal rod disk membranes from cattle have been examined by differential scanning calorimetry. Under conditions of 67 mM phosphate pH 7.0, unbleached rod outer segment disk membranes gave a single major endotherm with a temperature of denaturation (Tm) of 71.9 +/- 0.4 degrees C and a thermal unfolding calorimetric enthalpy change (delta Hcal) of 700 +/- 17 kJ/mol rhodopsin. Bleached rod outer segment disk membranes (membranes that had lost their absorbance at 498 nm after exposure to orange light) gave a single major endotherm with a Tm of 55.9 +/- 0.3 degrees C and a delta Hcal of 520 +/- 17 kJ/mol opsin. Neither bleached nor unbleached rod outer segment disk membranes gave endotherms upon thermal rescans. When thermal stability is examined over the pH range of 4-9, the major endotherms of both bleached and unbleached rod outer segment disk membranes were found to show maximum stability at pH 6.1. The observed delta Hcal values for bleached and unbleached rod outer segment disk membranes exhibit membrane concentration dependences which plateau at protein concentrations beyond 1.5 mg/mL. For partially bleached samples of rod outer segment disk membranes, the calorimetric enthalpy change for opsin appears to be somewhat dependent on the degree of bleaching, indicating intramembrane nearest neighbor interactions which affect the unfolding of opsin. Delta Hcal and Tm are particularly useful for assessing stability and testing for completeness of regeneration of rhodopsin from opsin. Other factors such as sample preparation and the presence of low concentrations of ethanol also affect the delta Hcal values while the Tm values remain fairly constant. This shows that the delta Hcal is a sensitive parameter for monitoring environmental changes of rhodopsin and opsin. Study holds ProTherm entries: 10025, 10026 Extra Details: additive : dithioerythritol(1 mM), retinal rod disk membranes; endotherm; membrane concentration;,nearest neighbor interactions

Submission Details

ID: kFDA5PuK4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:39 p.m.

Version: 1

Publication Details
Khan SM;Bolen W;Hargrave PA;Santoro MM;McDowell JH,Eur. J. Biochem. (1991) Differential scanning calorimetry of bovine rhodopsin in rod-outer-segment disk membranes. PMID:1831759
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
4UE5 2014-12-15T00:00:00+0000 9.0 Structural basis for targeting and elongation arrest of Bacillus signal recognition particle
1RY1 2003-12-19T00:00:00+0000 12.0 Structure of the signal recognition particle interacting with the elongation-arrested ribosome
2J28 2006-08-16T00:00:00+0000 8.0 MODEL OF E. COLI SRP BOUND TO 70S RNCS
4ZWJ 2015-05-19T00:00:00+0000 3.3 Crystal structure of rhodopsin bound to arrestin by femtosecond X-ray laser
5DGY 2015-08-28T00:00:00+0000 7.7 Crystal structure of rhodopsin bound to visual arrestin
5W0P 2017-05-31T00:00:00+0000 3.01 Crystal structure of rhodopsin bound to visual arrestin determined by X-ray free electron laser
5W0P 2017-05-31T00:00:00+0000 3.01 Crystal structure of rhodopsin bound to visual arrestin determined by X-ray free electron laser
6CMO 2018-03-05T00:00:00+0000 4.5 Rhodopsin-Gi complex
1EDS 2000-01-28T00:00:00+0000 0 SOLUTION STRUCTURE OF INTRADISKAL LOOP 1 OF BOVINE RHODOPSIN (RHODOPSIN RESIDUES 92-123)
1EDV 2000-01-28T00:00:00+0000 0 SOLUTION STRUCTURE OF 2ND INTRADISKAL LOOP OF BOVINE RHODOPSIN (RESIDUES 172-205)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.8 Rhodopsin Q68J47 OPSD_LOXAF
90.5 Rhodopsin O62793 OPSD_MESBI
92.2 Rhodopsin P51489 OPSD_RAT
92.2 Rhodopsin P28681 OPSD_CRIGR
91.7 Rhodopsin O62792 OPSD_GLOME
92.2 Rhodopsin P15409 OPSD_MOUSE
91.7 Rhodopsin O62798 OPSD_TURTR
92.0 Rhodopsin O62791 OPSD_DELDE
92.2 Rhodopsin O62795 OPSD_PAGGO
91.7 Rhodopsin O62796 OPSD_TRIMA
92.0 Rhodopsin P49912 OPSD_RABIT
92.2 Rhodopsin Q6W3E1 OPSD_CALPD
92.5 Rhodopsin O62794 OPSD_PHOVI
92.5 Rhodopsin P08100 OPSD_HUMAN
92.8 Rhodopsin Q769E8 OPSD_OTOCR
93.4 Rhodopsin P32308 OPSD_CANLF
93.7 Rhodopsin O18766 OPSD_PIG
94.3 Rhodopsin Q95KU1 OPSD_FELCA
95.4 Rhodopsin P02700 OPSD_SHEEP
100.0 Rhodopsin P02699 OPSD_BOVIN