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
1EDS 2000-08-09 SOLUTION STRUCTURE OF INTRADISKAL LOOP 1 OF BOVINE RHODOPSIN (RHODOPSIN RESIDUES 92-123)
1EDX 2000-08-09 SOLUTION STRUCTURE OF AMINO TERMINUS OF BOVINE RHODOPSIN (RESIDUES 1-40)
1EDV 2000-08-09 SOLUTION STRUCTURE OF 2ND INTRADISKAL LOOP OF BOVINE RHODOPSIN (RESIDUES 172-205)
1EDW 2000-08-09 SOLUTION STRUCTURE OF THIRD INTRADISKAL LOOP OF BOVINE RHODOPSIN (RESIDUES 268-293)
1FDF 2000-07-27 HELIX 7 BOVINE RHODOPSIN
1LN6 2002-07-10 STRUCTURE OF BOVINE RHODOPSIN (Metarhodopsin II)
1NZS 2004-03-02 NMR structures of phosphorylated carboxy terminus of bovine rhodopsin in arrestin-bound state
1JFP 2001-10-05 Structure of bovine rhodopsin (dark adapted)
1VQX 2005-01-18 ARRESTIN-BOUND NMR STRUCTURES OF THE PHOSPHORYLATED CARBOXY-TERMINAL DOMAIN OF RHODOPSIN, REFINED
1RY1 2004-04-20 12.0 Structure of the signal recognition particle interacting with the elongation-arrested ribosome
1U19 2004-10-12 2.2 Crystal Structure of Bovine Rhodopsin at 2.2 Angstroms Resolution
4X1H 2015-11-04 2.29 Opsin/G(alpha) peptide complex stabilized by nonyl-glucoside
5DYS 2016-08-10 2.3 Crystal Structure of T94I rhodopsin mutant
6FK6 2018-04-04 2.36 Crystal structure of N2C/D282C stabilized opsin bound to RS01
6FKC 2018-04-04 2.46 Crystal structure of N2C/D282C stabilized opsin bound to RS15
6FKD 2018-04-04 2.49 Crystal structure of N2C/D282C stabilized opsin bound to RS16
2G87 2006-09-02 2.6 Crystallographic model of bathorhodopsin
3OAX 2011-01-19 2.6 Crystal structure of bovine rhodopsin with beta-ionone
1L9H 2002-05-15 2.6 Crystal structure of bovine rhodopsin at 2.6 angstroms RESOLUTION
6FK7 2018-04-04 2.62 Crystal structure of N2C/D282C stabilized opsin bound to RS06
6FK9 2018-04-04 2.63 Crystal structure of N2C/D282C stabilized opsin bound to RS09
1GZM 2003-11-20 2.65 Structure of Bovine Rhodopsin in a Trigonal Crystal Form
4J4Q 2013-10-30 2.65 Crystal structure of active conformation of GPCR opsin stabilized by octylglucoside
3C9L 2008-08-05 2.65 Structure of ground-state bovine rhodospin in a hexagonal crystal form
6FKA 2018-04-04 2.7 Crystal structure of N2C/D282C stabilized opsin bound to RS11
5TE3 2017-03-15 2.7 Crystal structure of Bos taurus opsin at 2.7 Angstrom
4PXF 2014-09-17 2.75 Crystal structure of the active G-protein-coupled receptor opsin in complex with the finger-loop peptide derived from the full-length arrestin-1
2HPY 2006-08-22 2.8 Crystallographic model of lumirhodopsin
1HZX 2001-07-04 2.8 CRYSTAL STRUCTURE OF BOVINE RHODOPSIN
1F88 2000-08-04 2.8 CRYSTAL STRUCTURE OF BOVINE RHODOPSIN
5EN0 2016-08-10 2.81 Crystal Structure of T94I rhodopsin mutant
3PQR 2011-03-09 2.85 Crystal structure of Metarhodopsin II in complex with a C-terminal peptide derived from the Galpha subunit of transducin
6FK8 2018-04-04 2.87 Crystal structure of N2C/D282C stabilized opsin bound to RS08
4BEY 2013-05-08 2.9 Night blindness causing G90D rhodopsin in complex with GaCT2 peptide
3CAP 2008-06-24 2.9 Crystal Structure of Native Opsin: the G Protein-Coupled Receptor Rhodopsin in its Ligand-free State
2PED 2007-10-30 2.95 Crystallographic model of 9-cis-rhodopsin
3PXO 2011-03-09 3.0 Crystal structure of Metarhodopsin II
2X72 2011-03-16 3.0 CRYSTAL STRUCTURE OF THE CONSTITUTIVELY ACTIVE E113Q,D2C,D282C RHODOPSIN MUTANT WITH BOUND GALPHACT PEPTIDE.
5W0P 2017-08-09 3.01 Crystal structure of rhodopsin bound to visual arrestin determined by X-ray free electron laser
6FKB 2018-04-04 3.03 Crystal structure of N2C/D282C stabilized opsin bound to RS13
6FUF 2018-10-03 3.12 Crystal structure of the rhodopsin-mini-Go complex
5WKT 2017-12-13 3.2 3.2-Angstrom In situ Mylar structure of bovine opsin at 100 K
3DQB 2008-09-23 3.2 Crystal structure of the active G-protein-coupled receptor opsin in complex with a C-terminal peptide derived from the Galpha subunit of transducin
4ZWJ 2015-07-29 3.3 Crystal structure of rhodopsin bound to arrestin by femtosecond X-ray laser
6OYA 2019-07-24 3.3 Structure of the Rhodopsin-Transducin-Nanobody Complex
4A4M 2012-01-25 3.3 Crystal structure of the light-activated constitutively active N2C, M257Y,D282C rhodopsin mutant in complex with a peptide resembling the C-terminus of the Galpha-protein subunit (GaCT)
4BEZ 2013-04-24 3.3 Night blindness causing G90D rhodopsin in the active conformation
3C9M 2008-08-05 3.4 Structure of a mutant bovine rhodopsin in hexagonal crystal form
2J4Y 2007-09-25 3.4 Crystal structure of a rhodopsin stabilizing mutant expressed in mammalian cells
2I35 2006-10-17 3.8 Crystal structure of rhombohedral crystal form of ground-state rhodopsin
6OY9 2019-07-24 3.9 Structure of the Rhodopsin-Transducin Complex
5TE5 2017-03-15 4.01 Crystal structure of Bos taurus opsin regenerated with 6-carbon ring retinal chromophore
2I36 2006-10-17 4.1 Crystal structure of trigonal crystal form of ground-state rhodopsin
2I37 2006-10-17 4.15 Crystal structure of a photoactivated rhodopsin
6QNO 2019-07-10 4.38 Rhodopsin-Gi protein complex
6CMO 2018-06-20 4.5 Rhodopsin-Gi complex
5DGY 2016-03-23 7.7 Crystal structure of rhodopsin bound to visual arrestin
4UE5 2015-09-09 9.0 Structural basis for targeting and elongation arrest of Bacillus signal recognition particle
2J28 2006-11-08 9.5 MODEL OF E. COLI SRP BOUND TO 70S RNCS

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