Marco Capogna

Mutant alpha-latrotoxin (LTXN4C) does not form pores and causes secretion by receptor stimulation: this action does not require neurexins

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Standard

Mutant alpha-latrotoxin (LTXN4C) does not form pores and causes secretion by receptor stimulation : this action does not require neurexins. / Volynski, Kirill E; Capogna, Marco; Ashton, Anthony C et al.

In: The Journal of biological chemistry, Vol. 278, No. 33, 15.08.2003, p. 31058-66.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Harvard

Volynski, KE, Capogna, M, Ashton, AC, Thomson, D, Orlova, EV, Manser, CF, Ribchester, RR & Ushkaryov, YA 2003, 'Mutant alpha-latrotoxin (LTXN4C) does not form pores and causes secretion by receptor stimulation: this action does not require neurexins', The Journal of biological chemistry, vol. 278, no. 33, pp. 31058-66. https://doi.org/10.1074/jbc.M210395200

APA

Volynski, K. E., Capogna, M., Ashton, A. C., Thomson, D., Orlova, E. V., Manser, C. F., Ribchester, R. R., & Ushkaryov, Y. A. (2003). Mutant alpha-latrotoxin (LTXN4C) does not form pores and causes secretion by receptor stimulation: this action does not require neurexins. The Journal of biological chemistry, 278(33), 31058-66. https://doi.org/10.1074/jbc.M210395200

CBE

Volynski KE, Capogna M, Ashton AC, Thomson D, Orlova EV, Manser CF, Ribchester RR, Ushkaryov YA. 2003. Mutant alpha-latrotoxin (LTXN4C) does not form pores and causes secretion by receptor stimulation: this action does not require neurexins. The Journal of biological chemistry. 278(33):31058-66. https://doi.org/10.1074/jbc.M210395200

MLA

Vancouver

Volynski KE, Capogna M, Ashton AC, Thomson D, Orlova EV, Manser CF et al. Mutant alpha-latrotoxin (LTXN4C) does not form pores and causes secretion by receptor stimulation: this action does not require neurexins. The Journal of biological chemistry. 2003 Aug 15;278(33):31058-66. doi: 10.1074/jbc.M210395200

Author

Volynski, Kirill E ; Capogna, Marco ; Ashton, Anthony C et al. / Mutant alpha-latrotoxin (LTXN4C) does not form pores and causes secretion by receptor stimulation : this action does not require neurexins. In: The Journal of biological chemistry. 2003 ; Vol. 278, No. 33. pp. 31058-66.

Bibtex

@article{28e0cd34805d44519bcf815ede42cdac,
title = "Mutant alpha-latrotoxin (LTXN4C) does not form pores and causes secretion by receptor stimulation: this action does not require neurexins",
abstract = "Alpha-latrotoxin (LTX) causes massive release of neurotransmitters via a complex mechanism involving (i) activation of receptor(s) and (ii) toxin insertion into the plasma membrane with (iii) subsequent pore formation. Using cryo-electron microscopy, electrophysiological and biochemical methods, we demonstrate here that the recently described toxin mutant (LTXN4C) is unable to insert into membranes and form pores due to its inability to assemble into tetramers. However, this mutant still binds to major LTX receptors (latrophilin and neurexin) and causes strong transmitter exocytosis in synaptosomes, hippocampal slice cultures, neuromuscular junctions, and chromaffin cells. In the absence of mutant incorporation into the membrane, receptor activation must be the only mechanism by which LTXN4C triggers exocytosis. An interesting feature of this receptor-mediated transmitter release is its dependence on extracellular Ca2+. Because Ca2+ is also strictly required for LTX interaction with neurexin, the latter might be the only receptor mediating the LTXN4C action. To test this hypothesis, we used conditions (substitution of Ca2+ in the medium with Sr2+) under which LTXN4C does not bind to any member of the neurexin family but still interacts with latrophilin. We show that, in all the systems tested, Sr2+ fully replaces Ca2+ in supporting the stimulatory effect of LTXN4C. These results indicate that LTXN4C can cause neurotransmitter release just by stimulating a receptor and that neurexins are not critical for this receptor-mediated action.",
keywords = "Acetylcholine/metabolism, Animals, Black Widow Spider, CHO Cells, COS Cells, Calcium/metabolism, Carbon Radioisotopes, Catecholamines/metabolism, Cattle, Cell Membrane/metabolism, Chromaffin Cells/metabolism, Cricetinae, Exocytosis, Glutamic Acid/pharmacokinetics, Hippocampus/cytology, Membrane Potentials/physiology, Mice, Mutation, Neuromuscular Junction/metabolism, Patch-Clamp Techniques, Protein Structure, Quaternary, Rats, Receptors, Cell Surface/metabolism, Spider Venoms/chemistry, Strontium/pharmacology, Synaptosomes/metabolism",
author = "Volynski, {Kirill E} and Marco Capogna and Ashton, {Anthony C} and Derek Thomson and Orlova, {Elena V} and Manser, {Catherine F} and Ribchester, {Richard R} and Ushkaryov, {Yuri A}",
year = "2003",
month = aug,
day = "15",
doi = "10.1074/jbc.M210395200",
language = "English",
volume = "278",
pages = "31058--66",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "33",

}

RIS

TY - JOUR

T1 - Mutant alpha-latrotoxin (LTXN4C) does not form pores and causes secretion by receptor stimulation

T2 - this action does not require neurexins

AU - Volynski, Kirill E

AU - Capogna, Marco

AU - Ashton, Anthony C

AU - Thomson, Derek

AU - Orlova, Elena V

AU - Manser, Catherine F

AU - Ribchester, Richard R

AU - Ushkaryov, Yuri A

PY - 2003/8/15

Y1 - 2003/8/15

N2 - Alpha-latrotoxin (LTX) causes massive release of neurotransmitters via a complex mechanism involving (i) activation of receptor(s) and (ii) toxin insertion into the plasma membrane with (iii) subsequent pore formation. Using cryo-electron microscopy, electrophysiological and biochemical methods, we demonstrate here that the recently described toxin mutant (LTXN4C) is unable to insert into membranes and form pores due to its inability to assemble into tetramers. However, this mutant still binds to major LTX receptors (latrophilin and neurexin) and causes strong transmitter exocytosis in synaptosomes, hippocampal slice cultures, neuromuscular junctions, and chromaffin cells. In the absence of mutant incorporation into the membrane, receptor activation must be the only mechanism by which LTXN4C triggers exocytosis. An interesting feature of this receptor-mediated transmitter release is its dependence on extracellular Ca2+. Because Ca2+ is also strictly required for LTX interaction with neurexin, the latter might be the only receptor mediating the LTXN4C action. To test this hypothesis, we used conditions (substitution of Ca2+ in the medium with Sr2+) under which LTXN4C does not bind to any member of the neurexin family but still interacts with latrophilin. We show that, in all the systems tested, Sr2+ fully replaces Ca2+ in supporting the stimulatory effect of LTXN4C. These results indicate that LTXN4C can cause neurotransmitter release just by stimulating a receptor and that neurexins are not critical for this receptor-mediated action.

AB - Alpha-latrotoxin (LTX) causes massive release of neurotransmitters via a complex mechanism involving (i) activation of receptor(s) and (ii) toxin insertion into the plasma membrane with (iii) subsequent pore formation. Using cryo-electron microscopy, electrophysiological and biochemical methods, we demonstrate here that the recently described toxin mutant (LTXN4C) is unable to insert into membranes and form pores due to its inability to assemble into tetramers. However, this mutant still binds to major LTX receptors (latrophilin and neurexin) and causes strong transmitter exocytosis in synaptosomes, hippocampal slice cultures, neuromuscular junctions, and chromaffin cells. In the absence of mutant incorporation into the membrane, receptor activation must be the only mechanism by which LTXN4C triggers exocytosis. An interesting feature of this receptor-mediated transmitter release is its dependence on extracellular Ca2+. Because Ca2+ is also strictly required for LTX interaction with neurexin, the latter might be the only receptor mediating the LTXN4C action. To test this hypothesis, we used conditions (substitution of Ca2+ in the medium with Sr2+) under which LTXN4C does not bind to any member of the neurexin family but still interacts with latrophilin. We show that, in all the systems tested, Sr2+ fully replaces Ca2+ in supporting the stimulatory effect of LTXN4C. These results indicate that LTXN4C can cause neurotransmitter release just by stimulating a receptor and that neurexins are not critical for this receptor-mediated action.

KW - Acetylcholine/metabolism

KW - Animals

KW - Black Widow Spider

KW - CHO Cells

KW - COS Cells

KW - Calcium/metabolism

KW - Carbon Radioisotopes

KW - Catecholamines/metabolism

KW - Cattle

KW - Cell Membrane/metabolism

KW - Chromaffin Cells/metabolism

KW - Cricetinae

KW - Exocytosis

KW - Glutamic Acid/pharmacokinetics

KW - Hippocampus/cytology

KW - Membrane Potentials/physiology

KW - Mice

KW - Mutation

KW - Neuromuscular Junction/metabolism

KW - Patch-Clamp Techniques

KW - Protein Structure, Quaternary

KW - Rats

KW - Receptors, Cell Surface/metabolism

KW - Spider Venoms/chemistry

KW - Strontium/pharmacology

KW - Synaptosomes/metabolism

U2 - 10.1074/jbc.M210395200

DO - 10.1074/jbc.M210395200

M3 - Journal article

C2 - 12782639

VL - 278

SP - 31058

EP - 31066

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 33

ER -