Fluorescent- and tagged-protoxin II peptides: potent markers of the Nav1.7 channel pain target

被引:8
作者
Montnach, Jerome [1 ]
De Waard, Stephan [1 ]
Nicolas, Sebastien [1 ]
Burel, Sophie [1 ]
Osorio, Nancy [2 ]
Zoukimian, Claude [3 ]
Mantegazza, Massimo [4 ]
Boukaiba, Rachid [5 ]
Beroud, Remy [3 ]
Partiseti, Michel [5 ]
Delmas, Patrick [2 ]
Marionneau, Celine [1 ]
De Waard, Michel [1 ,3 ]
机构
[1] Univ Nantes, CNRS, INSERM, LabEx Ion Channels Sci & Therapeut,Inst Thorax, Nantes, France
[2] Aix Marseille Univ, CNRS, UMR 7291, Lab Cognit Neurosci, Marseille, France
[3] Smartox Biotechnol, St Egreve, France
[4] Univ Cote Azur, CNRS, UMR 7275, Inst Mol & Cellular Pharmacol, Valbonne, France
[5] Sanofi R&D, Integrated Drug Discovery High Content Biol, Vitry Sur Seine, France
关键词
automated patch‐ clamp; biotinylated analogue; cell line; cellular distribution; fluorescent analogue; Na(v)1; 7; pain target; protoxin II; pull‐ down; voltage‐ gated sodium channel; PROTX-II; SODIUM-CHANNELS; CELL PENETRATION; VOLTAGE SENSOR; MAUROCALCINE; CONTRIBUTES; MUTATIONS; PHARMACOLOGY; ACTIVATION; DESIGN;
D O I
10.1111/bph.15453
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Background and Purpose Protoxin II (ProTx II) is a high affinity gating modifier that is thought to selectively block the Na(v)1.7 voltage-dependent Na+ channel, a major therapeutic target for the control of pain. We aimed at producing ProTx II analogues entitled with novel functionalities for cell distribution studies and biochemical characterization of its Na-v channel targets. Experimental Approach We took advantage of the high affinity properties of the peptide, combined to its slow off rate, to design a number of new tagged analogues useful for imaging and biochemistry purposes. We used high-throughput automated patch-clamp to identify the analogues best matching the native properties of ProTx II and validated them on various Na-v-expressing cells in pull-down and cell distribution studies. Key Results Two of the produced ProTx II analogues, Biot-ProTx II and ATTO488-ProTx II, best emulate the pharmacological properties of unlabelled ProTx II, whereas other analogues remain high affinity blockers of Na(v)1.7. The biotinylated version of ProTx II efficiently works for the pull-down of several Na-v isoforms tested in a concentration-dependent manner, whereas the fluorescent ATTO488-ProTx II specifically labels the Na(v)1.7 channel over other Na-v isoforms tested in various experimental conditions. Conclusions and Implications The properties of these ProTx II analogues as tools for Na-v channel purification and cell distribution studies pave the way for a better understanding of ProTx II channel receptors in pain and their pathophysiological implications in sensory neuronal processing. The new fluorescent ProTx II should also be useful in the design of new drug screening strategies.
引用
收藏
页码:2632 / 2650
页数:19
相关论文
共 41 条
[1]   Building sensory axons: Delivery and distribution of Nav1.7 channels and effects of inflammatory mediators [J].
Akin, Elizabeth J. ;
Higerd, Grant P. ;
Mis, Malgorzata A. ;
Tanaka, Brian S. ;
Adi, Talia ;
Liu, Shujun ;
Dib-Hajj, Fadia B. ;
Waxman, Stephen G. ;
Dib-Hajj, Sulayman D. .
SCIENCE ADVANCES, 2019, 5 (10)
[2]   THE CONCISE GUIDE TO PHARMACOLOGY 2019/20: Ion channels [J].
Alexander, Stephen P. H. ;
Mathie, Alistair ;
Peters, John A. ;
Veale, Emma L. ;
Striessnig, Joerg ;
Kelly, Eamonn ;
Armstrong, Jane F. ;
Faccenda, Elena ;
Harding, Simon D. ;
Pawson, Adam J. ;
Sharman, Joanna L. ;
Southan, Christopher ;
Davies, Jamie A. .
BRITISH JOURNAL OF PHARMACOLOGY, 2019, 176 :S142-S228
[3]  
Bilimoria P.M, 2008, CSH PROTOC 2008 PDB
[4]   Cell penetration properties of maurocalcine, a natural venom peptide active on the intracellular ryanodine receptor [J].
Boisseau, Sylvie ;
Mabrouk, Kamel ;
Ram, Narendra ;
Garmy, Nicolas ;
Collin, Veronique ;
Tadmouri, Abir ;
Mikati, Mohamad ;
Sabatier, Jean-Marc ;
Ronjat, Michel ;
Fantini, Jacques ;
De Waard, Michel .
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 2006, 1758 (03) :308-319
[5]   Voltage-gated sodium channels assemble and gate as dimers [J].
Clatot, Jerome ;
Hoshi, Malcolm ;
Wan, Xiaoping ;
Liu, Haiyan ;
Jain, Ankur ;
Shinlapawittayatorn, Krekwit ;
Marionneau, Celine ;
Ficker, Eckhard ;
Ha, Taekjip ;
Deschenes, Isabelle .
NATURE COMMUNICATIONS, 2017, 8
[6]   Pharmacological dissection and distribution of NaN/Nav1.9, T-type Ca2+ currents, and mechanically activated cation currents in different populations of DRG neurons [J].
Coste, Bertrand ;
Crest, Marcel ;
Delmas, Patrick .
JOURNAL OF GENERAL PHYSIOLOGY, 2007, 129 (01) :57-77
[7]   An SCN9A channelopathy causes congenital inability to experience pain [J].
Cox, James J. ;
Reimann, Frank ;
Nicholas, Adeline K. ;
Thornton, Gemma ;
Roberts, Emma ;
Springell, Kelly ;
Karbani, Gulshan ;
Jafri, Hussain ;
Mannan, Jovaria ;
Raashid, Yasmin ;
Al-Gazali, Lihadh ;
Hamamy, Henan ;
Valente, Enza Maria ;
Gorman, Shaun ;
Williams, Richard ;
McHale, Duncan P. ;
Wood, John N. ;
Gribble, Fiona M. ;
Woods, C. Geoffrey .
NATURE, 2006, 444 (7121) :894-898
[8]   Electrophysiological properties of mutant Nav1.7 sodium channels in a painful inherited neuropathy [J].
Cummins, TR ;
Dib-Hajj, SD ;
Waxman, SG .
JOURNAL OF NEUROSCIENCE, 2004, 24 (38) :8232-8236
[9]   Experimental design and analysis and their reporting II: updated and simplified guidance for authors and peer reviewers [J].
Curtis, Michael J. ;
Alexander, Steve ;
Cirino, Giuseppe ;
Docherty, James R. ;
George, Christopher H. ;
Giembycz, Mark A. ;
Hoyer, Daniel ;
Insel, Paul A. ;
Izzo, Angelo A. ;
Ji, Yong ;
MacEwan, David J. ;
Sobey, Christopher G. ;
Stanford, S. Clare ;
Teixeira, Mauro M. ;
Wonnacott, Sue ;
Ahluwalia, Amrita .
BRITISH JOURNAL OF PHARMACOLOGY, 2018, 175 (07) :987-993
[10]   Gain-of-function mutation in Nav1.7 in familial erythromelalgia induces bursting of sensory neurons [J].
Dib-Hajj, SD ;
Rush, AM ;
Cummins, TR ;
Hisama, FM ;
Novella, S ;
Tyrrell, L ;
Marshall, L ;
Waxman, SG .
BRAIN, 2005, 128 :1847-1854