Structural domains required for channel function of the mouse transient receptor potential protein homologue TRP1β

FEBS Letters

Budde, Petra²  Friedrich, Olaf²  Jüngling, Eberhard¹  Rocks, Oliver³  Engelke, Michael²  Zitt, Christof¹  Frey, Jürgen²  Lückhoff, Andreas¹  Schäfer, Christina²  Niemann, Ursula² 

[1] Institut für Physiologie, RWTH-Aachen, Pauwelsstrasse 30, D-52074 Aachen, Germany;Universität Bielefeld, Fakultät für Chemie, Biochemie II, Universitätsstrasse 25, D-33615 Bielefeld, Germany;Max-Planck-Institut für Molekulare Physiologie, Otto-Hahn-Str. 11, D-44227 Dortmund, Germany

关键词: Transient receptor potential protein;    Coiled-coil domain;    Ankyrin-like repeat;    HEK293 cell;    Patch clamp;    Yeast two-hybrid system;    ANK-repeat;    ankyrin-like repeat;    [Ca2+]i;    intracellular free calcium concentration;    CCh;    carbachol;    cc;    coiled-coil;    ECS;    extracellular solution;    EGFP;    enhanced green fluorescent protein;    fura-2-AM;    fura-2-acetoxymethyl ester;    HEK293;    human embryonic kidney cells;    IP3;    inositol-1;    4;    5-trisphosphate;    mAb;    monoclonal antibody;    M r;    relative molecular mass;    mTRP;    mouse TRP;    TRP;    transient receptor potential protein;    NMDG;    N-methyl-D-glucamine;    pp;    putative pore;    TRPC;    TRP channel;   

DOI  :  10.1016/S0014-5793(02)02971-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Transient receptor potential proteins (TRP) are supposed to participate in the formation of store-operated Ca²⁺ influx channels by co-assembly. However, little is known which domains facilitate the interaction of subunits. Contribution of the N-terminal coiled-coil domain and ankyrin-like repeats and the putative pore region of the mouse TRP1β (mTRP1β) variant to the formation of functional cation channels were analyzed following overexpression in HEK293 (human embryonic kidney) cells. MTRP1β expressing cells exhibited enhanced Ca²⁺ influx and enhanced whole-cell membrane currents compared to mTRP1β deletion mutants. Using a yeast two-hybrid assay only the coiled-coil domain facilitated homodimerization of the N-terminus. These results suggest that the N-terminus of mTRP1β is required for structural organization thus forming functional channels.

【 授权许可】

Unknown