Structure and function in rhodopsin: Mass spectrometric identification of the abnormal intradiscal disulfide bond in misfolded retinitis pigmentosa mutants

被引：54
作者：
Hwa, J
Klein-Seetharaman, J
Khorana, HG
机构：
[1] MIT, Dept Biol, Cambridge, MA 02139 USA
[2] MIT, Dept Chem, Cambridge, MA 02139 USA
来源：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2001年 / 98卷 / 09期
关键词：
G protein-coupled receptors; signal transduction; transmembrane domain; N-ethylmaleimide; biotin-avidin affinity chromatography;
D O I：
10.1073/pnas.061632798
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
Retinitis pigmentosa (RP) point mutations in both the intradiscal (ID) and transmembrane domains of rhodopsin cause partial or complete misfolding of rhodopsin. resulting in loss of Il-cis-retinal binding. Previous work has shown that misfolding is caused by the formation of a disulfide bond in the ID domain different from the native Cys-110-Cys-187 disulfide bond in native rhodopsin. Here we report on direct identification of the abnormal disulfide bond in misfolded RP mutants in the transmembrane domain by mass spectrometric analysis. This disulfide bond is between Cys-185 and Cys-187. the same as previously identified in misfolded RP mutations in the ID domain. The strategy described here should be generally applicable to identification of disulfide bonds in other integral membrane proteins.
引用
收藏
页码：4872 / 4876
页数：5
相关论文
共 26 条
[1]   ON THE DISULFIDE BONDS OF RHODOPSINS [J].
ALSALEH, S ;
GORE, M ;
AKHTAR, M .
BIOCHEMICAL JOURNAL, 1987, 246 (01) :131-137
[2]  
ANUKANTH A, 1994, J BIOL CHEM, V269, P19738
[3]   An alpha-carbon template for the transmembrane helices in the rhodopsin family of G-protein-coupled receptors [J].
Baldwin, JM ;
Schertler, GFX ;
Unger, VM .
JOURNAL OF MOLECULAR BIOLOGY, 1997, 272 (01) :144-164
[4]  
BERSON EL, 1993, INVEST OPHTH VIS SCI, V34, P1659
[5]   LOCATION OF DISULPHIDE BRIDGES BY DIAGONAL PAPER ELECTROPHORESIS - DISULPHIDE BRIDGES OF BOVINE CHYMOTRYPSINOGEN A [J].
BROWN, JR ;
HARTLEY, BS .
BIOCHEMICAL JOURNAL, 1966, 101 (01) :214-&
[6]   Structure and function in rhodopsin: Destabilization of rhodopsin by the binding of an antibody at the N-terminal segment provides support for involvement of the latter in an intradiscal tertiary structure [J].
Cha, KW ;
Reeves, PJ ;
Khorana, HG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (07) :3016-3021
[7]  
DOI T, 1990, P NATL ACAD SCI USA, V87, P4991, DOI 10.1073/pnas.87.13.4991
[8]   Structure and function in rhodopsin: Correct folding and misfolding in point mutants at and in proximity to the site of the retinitis pigmentosa mutation Leu-125->Arg in the transmembrane helix C [J].
Garriga, P ;
Liu, X ;
Khorana, HG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (10) :4560-4564
[9]   DETERMINATION OF SULFHYDRYL GROUPS WITH 2,2'- OR 4,4'-DITHIODIPYRIDINE [J].
GRASSETTI, DR ;
MURRAY, JF .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1967, 119 (1-3) :41-+
[10]   Structure and function in rhodopsin: Packing of the helices in the transmembrane domain and folding to a tertiary structure in the intradiscal domain are coupled [J].
Hwa, J ;
Garriga, P ;
Liu, X ;
Khorana, HG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (20) :10571-10576
123