TY  - JOUR
A1  - Nicolas, Sébastien
A1  - Zoukimian, Claude
A1  - Bosmans, Frank
A1  - Montnach, Jérôme
A1  - Diochot, Sylvie
A1  - Cuypers, Eva
A1  - Waard, Stephan de
A1  - Béroud, Rémy
A1  - Mebs, Dietrich
A1  - Craik, David
A1  - Boturyn, Didier
A1  - Lazdunski, Michel
A1  - Tytgat, Jan
A1  - Waard, Michel de
T1  - Chemical synthesis, proper folding, nav channel selectivity profile and analgesic properties of the spider peptide phlotoxin 1
T2  - Toxins
N2  - Phlotoxin-1 (PhlTx1) is a peptide previously identified in tarantula venom (Phlogius species) that belongs to the inhibitory cysteine-knot (ICK) toxin family. Like many ICK-based spider toxins, the synthesis of PhlTx1 appears particularly challenging, mostly for obtaining appropriate folding and concomitant suitable disulfide bridge formation. Herein, we describe a procedure for the chemical synthesis and the directed sequential disulfide bridge formation of PhlTx1 that allows for a straightforward production of this challenging peptide. We also performed extensive functional testing of PhlTx1 on 31 ion channel types and identified the voltage-gated sodium (Nav) channel Nav1.7 as the main target of this toxin. Moreover, we compared PhlTx1 activity to 10 other spider toxin activities on an automated patch-clamp system with Chinese Hamster Ovary (CHO) cells expressing human Nav1.7. Performing these analyses in reproducible conditions allowed for classification according to the potency of the best natural Nav1.7 peptide blockers. Finally, subsequent in vivo testing revealed that intrathecal injection of PhlTx1 reduces the response of mice to formalin in both the acute pain and inflammation phase without signs of neurotoxicity. PhlTx1 is thus an interesting toxin to investigate Nav1.7 involvement in cellular excitability and pain
KW  - spider toxin
KW  - directed disulfide bond formation
KW  - Nav channel activity
KW  - Nav1.7
KW  - pain target
KW  - automated patch-clamp
Y1  - 2019
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/53532
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-535325
SN  - 2072-6651
VL  - 11
IS  - 367
PB  - MDPI
CY  - Basel
ER  -