Abstract
Transient receptor potential (TRP) channels as cellular sensors are thought to function as tetramers. Yet, the molecular determinants governing channel multimerization remain largely elusive. Here we report the identification of a segment comprising21amino acids(residues752-772 of mouse TRPV1) after the known TRP-like domaininthe channel Cterminus that functions as a tetrameric assembly domain (TAD). Purified recombinant C-terminal proteins of TRPV1-4, but not the N terminus, mediated the protein-protein interaction in an in vitro pulldown assay. Western blot analysis combined with electrophysiology and calcium imaging demonstrated that TAD exerted a robust dominant-negative effect on wild-type TRPV1. When fused with the membrane-tethered peptide Gap43, the TAD blocked the formation of stable homomultimers. Calcium imaging and current recordings showed that deletion of the TAD in a poreless TRPV1 mutant subunit suppressed its dominant-negative phenotype, confirming the involvement of the TAD in assembly of functional channels. Our findings suggest that the C-terminal TAD in TRPV1 channels functions as a domain that is conserved among TRPV1-4 and mediates a direct subunit-subunit interaction for tetrameric assembly.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 15308-15316 |
| Number of pages | 9 |
| Journal | Journal of Biological Chemistry |
| Volume | 286 |
| Issue number | 17 |
| DOIs | |
| State | Published - Apr 29 2011 |
Fingerprint
ASJC Scopus subject areas
- Biochemistry
- Cell Biology
- Molecular Biology
Cite this
Identification of a tetrameric assembly domain in the C terminus of heat-activated TRPV1 channels. / Zhang, Feng; Liu, Shuang; Yang, Fan; Zheng, Jie; Wang, Kewei.
In: Journal of Biological Chemistry, Vol. 286, No. 17, 29.04.2011, p. 15308-15316.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Identification of a tetrameric assembly domain in the C terminus of heat-activated TRPV1 channels
AU - Zhang, Feng
AU - Liu, Shuang
AU - Yang, Fan
AU - Zheng, Jie
AU - Wang, Kewei
PY - 2011/4/29
Y1 - 2011/4/29
N2 - Transient receptor potential (TRP) channels as cellular sensors are thought to function as tetramers. Yet, the molecular determinants governing channel multimerization remain largely elusive. Here we report the identification of a segment comprising21amino acids(residues752-772 of mouse TRPV1) after the known TRP-like domaininthe channel Cterminus that functions as a tetrameric assembly domain (TAD). Purified recombinant C-terminal proteins of TRPV1-4, but not the N terminus, mediated the protein-protein interaction in an in vitro pulldown assay. Western blot analysis combined with electrophysiology and calcium imaging demonstrated that TAD exerted a robust dominant-negative effect on wild-type TRPV1. When fused with the membrane-tethered peptide Gap43, the TAD blocked the formation of stable homomultimers. Calcium imaging and current recordings showed that deletion of the TAD in a poreless TRPV1 mutant subunit suppressed its dominant-negative phenotype, confirming the involvement of the TAD in assembly of functional channels. Our findings suggest that the C-terminal TAD in TRPV1 channels functions as a domain that is conserved among TRPV1-4 and mediates a direct subunit-subunit interaction for tetrameric assembly.
AB - Transient receptor potential (TRP) channels as cellular sensors are thought to function as tetramers. Yet, the molecular determinants governing channel multimerization remain largely elusive. Here we report the identification of a segment comprising21amino acids(residues752-772 of mouse TRPV1) after the known TRP-like domaininthe channel Cterminus that functions as a tetrameric assembly domain (TAD). Purified recombinant C-terminal proteins of TRPV1-4, but not the N terminus, mediated the protein-protein interaction in an in vitro pulldown assay. Western blot analysis combined with electrophysiology and calcium imaging demonstrated that TAD exerted a robust dominant-negative effect on wild-type TRPV1. When fused with the membrane-tethered peptide Gap43, the TAD blocked the formation of stable homomultimers. Calcium imaging and current recordings showed that deletion of the TAD in a poreless TRPV1 mutant subunit suppressed its dominant-negative phenotype, confirming the involvement of the TAD in assembly of functional channels. Our findings suggest that the C-terminal TAD in TRPV1 channels functions as a domain that is conserved among TRPV1-4 and mediates a direct subunit-subunit interaction for tetrameric assembly.
UR - http://www.scopus.com/inward/record.url?scp=79955439687&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79955439687&partnerID=8YFLogxK
U2 - 10.1074/jbc.M111.223941
DO - 10.1074/jbc.M111.223941
M3 - Article
C2 - 21357419
AN - SCOPUS:79955439687
VL - 286
SP - 15308
EP - 15316
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 17
ER -