Structural requirements for α-latrotoxin binding and α-latrotoxin-stimulated secretion -: A study with calcium-independent receptor of α-latrotoxin (CIRL) deletion mutants

被引:67
作者
Krasnoperov, V
Bittner, MA
Holz, RW
Chepurny, O
Petrenko, AG
机构
[1] NYU Med Ctr, Dept Pharmacol, New York, NY 10016 USA
[2] NYU Med Ctr, Dept Physiol & Neurosci, New York, NY 10016 USA
[3] NYU Med Ctr, Dept Environm Med, New York, NY 10016 USA
[4] Univ Michigan, Sch Med, Dept Pharmacol, Ann Arbor, MI 48109 USA
关键词
D O I
10.1074/jbc.274.6.3590
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
Stimulation of neurotransmitter release by alpha-latrotoxin requires its binding to the calcium-independent receptor of alpha-latrotoxin (CIRL), an orphan neuronal G protein-coupled receptor. CIRL consists of two noncovalently bound subunits, p85, a heptahelical integral membrane protein, and p120, a large extracellular polypeptide with domains homologous to lectin, olfactomedin, mucin, the secretin receptor family, and a novel structural motif common for large orphan G protein-coupled receptors, The analysis of CIRL deletion mutants indicates that the high affinity alpha-latrotoxin-binding site is located within residues 467-891, which comprise the first transmembrane segment of p85 and the C-terminal half of p120, The N-terminal lectin, olfactomedin, and mucin domains of p120 are not required for the interaction with alpha-latrotoxin. Soluble p120 and all its fragments, which include the 467-770 residues, bind alpha-latrotoxin with low affinity suggesting the importance of membrane-embedded p85 for the stabilization of the complex of the toxin with p120, Two COOH-terminal deletion mutants of CIRL, one with the truncated cytoplasmic domain and the other with only one transmembrane segment left of seven, supported both alpha-latrotoxin-induced calcium uptake in HEK293 cells and alpha-latrotoxin-stimulated secretion when expressed in chromaffin cells, although with a different dose dependence than wild-type CIRL and its N-terminal deletion mutant, Thus the signaling domains of CIRL are not critically important for the stimulation of exocytosis in intact chromaffin cells by alpha-latrotoxin.
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页码:3590 / 3596
页数:7
相关论文
共 20 条
[1]  
Bittner MA, 1998, J NEUROSCI, V18, P2914
[2]   Calcium-independent actions of alpha-latrotoxin on spontaneous and evoked synaptic transmission in the hippocampus [J].
Capogna, M ;
Gahwiler, BH ;
Thompson, SM .
JOURNAL OF NEUROPHYSIOLOGY, 1996, 76 (05) :3149-3158
[3]   Vesicle exocytosis stimulated by α-latrotoxin is mediated by latrophilin and requires both external and stored Ca2+ [J].
Davletov, BA ;
Meunier, FA ;
Ashton, AC ;
Matsushita, H ;
Hirst, WD ;
Lelianova, VG ;
Wilkin, GP ;
Dolly, JO ;
Ushkaryov, YA .
EMBO JOURNAL, 1998, 17 (14) :3909-3920
[4]   Isolation and biochemical characterization of a Ca2+-independent alpha-latrotoxin-binding protein [J].
Davletov, BA ;
Shamotienko, OG ;
Lelianova, VG ;
Grishin, EV ;
Ushkaryov, YA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (38) :23239-23245
[5]  
DOHLMAN HG, 1991, ANNU REV BIOCHEM, V60, P653, DOI 10.1146/annurev.biochem.60.1.653
[6]   Neurexin Iα is a major α-latratoxin receptor that cooperates in α-latrotoxin action [J].
Geppert, M ;
Khvotchev, M ;
Krasnoperov, V ;
Goda, Y ;
Missler, M ;
Hammer, RE ;
Ichtchenko, K ;
Petrenko, AG ;
Südhof, TC .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (03) :1705-1710
[7]  
ICHTCHENKO K, 1999, IN PRESS J BIOL CHEM
[8]   The calcium-independent receptor of alpha-latrotoxin is not a neurexin [J].
Krasnoperov, VG ;
Beavis, R ;
Chepurny, OG ;
Little, AR ;
Plotnikov, AN ;
Petrenko, AG .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1996, 227 (03) :868-875
[9]   alpha-latrotoxin stimulates exocytosis by the interaction with a neuronal G-protein-coupled receptor [J].
Krasnoperov, VG ;
Bittner, MA ;
Beavis, R ;
Kuang, YN ;
Salnikow, KV ;
Chepurny, OG ;
Little, AR ;
Plotnikov, AN ;
Wu, DQ ;
Holz, RW ;
Petrenko, AG .
NEURON, 1997, 18 (06) :925-937
[10]   Ca2+-independent insulin exocytosis induced by α-latrotoxin requires latrophilin, a G protein-coupled receptor [J].
Lang, JC ;
Ushkaryov, Y ;
Grasso, A ;
Wollheim, CB .
EMBO JOURNAL, 1998, 17 (03) :648-657