The Pseudo Signal Peptide of the Corticotropin-releasing Factor Receptor Type 2A Prevents Receptor Oligomerization

被引:27
作者
Teichmann, Anke [1 ]
Rutz, Claudia [1 ]
Kreuchwig, Annika [1 ]
Krause, Gerd [1 ]
Wiesner, Burkhard [1 ]
Schuelein, Ralf [1 ]
机构
[1] Leibniz Inst Mol Pharmacol, D-13125 Berlin, Germany
关键词
FLUORESCENCE CORRELATION SPECTROSCOPY; DELTA-OPIOID RECEPTORS; HORMONE-RECEPTOR; TRANSMEMBRANE DOMAIN; PROTEIN; DIMERIZATION; EXPRESSION; VARIANT; STRESS; CELL;
D O I
10.1074/jbc.M112.360594
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
N-terminal signal peptides mediate the interaction of native proteins with the translocon complex of the endoplasmic reticulum membrane and are cleaved off during early protein biogenesis. The corticotropin-releasing factor receptor type 2a (CRF2(a)R) possesses an N-terminal pseudo signal peptide, which represents a so far unique domain within the large protein family of G protein-coupled receptors (GPCRs). In contrast to a conventional signal peptide, the pseudo signal peptide remains uncleaved and consequently forms a hydrophobic extension at the N terminus of the receptor. The functional consequence of the presence of the pseudo signal peptide is not understood. Here, we have analyzed the significance of this domain for receptor dimerization/oligomerization in detail. To this end, we took the CRF2(a)R and the homologous corticotropin-releasing factor receptor type 1 (CRF1R) possessing a conventional cleaved signal peptide and conducted signal peptide exchange experiments. Using single cell and single molecule imaging methods (fluorescence resonance energy transfer and fluorescence cross-correlation spectroscopy, respectively) as well as biochemical experiments, we obtained two novel findings; we could show that (i) the CRF2(a)R is expressed exclusively as a monomer, and (ii) the presence of the pseudo signal peptide prevents its oligomerization. Thus, we have identified a novel functional domain within the GPCR protein family, which plays a role in receptor oligomerization and which may be useful to study the functional significance of this process in general.
引用
收藏
页码:27265 / 27274
页数:10
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