Expression of a functional recombinant Phoneutria nigriventer toxin active on K+ channels

被引：15

作者：

Carneiro, AMD

Kushmerick, C

Koenen, J

Arndt, MHL

Cordeiro, MN

Chavez-Olortegui, C

Diniz, CR

Gomez, MV

Kalapothakis, E

Prado, MAM

Prado, VF

机构：

[1] Univ Fed Minas Gerais, ICB, Dept Bioquim Imunol, Mol Neurobiol Lab, BR-31270901 Belo Horizonte, MG, Brazil

[2] Univ Fed Minas Gerais, ICB, Dept Farmacol, BR-31270901 Belo Horizonte, MG, Brazil

来源：

TOXICON | 2003年 / 41卷 / 03期

关键词：

spider toxins; K+ channels; GH3; cells; Phoneutria nigriventer;

D O I：

10.1016/S0041-0101(02)00292-1

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

PnTx3-1 is a peptide isolated from the venom of the spider Phoneutria nigriventer that specifically inhibits A-type K+ currents (I-A) in GH(3) cells. Here we used a bacterial expression system to produce an NH2-extended mutant of PnTx3-1 (ISEF-PnTx3-1) and tested whether the toxin is functional. The recombinant toxin was purified from bacterial extracts by a combination of affinity and ion-exchange chromatography. The recombinant toxin blocked A-type K+ currents in GH3 cells in a fashion similar to that observed with the wild-type toxin purified from the spider venom. These results suggest that recombinant cDNA methods provide a novel source for the production of functional Phoneutria toxins. The recombinant ISEF-PnTx3-1 should be useful for further understanding of the role of A-type K+ currents in biological processes. (C) 2003 Elsevier Science Ltd. All rights reserved.

引用

页码：305 / 313

页数：9

共 51 条

[1] TOPOLOGY OF THE PORE-REGION OF A K+ CHANNEL REVEALED BY THE NMR-DERIVED STRUCTURES OF SCORPION TOXINS [J].

AIYAR, J ;

WITHKA, JM ;

RIZZI, JP ;

SINGLETON, DH ;

ANDREWS, GC ;

LIN, W ;

BOYD, J ;

HANSON, DC ;

SIMON, M ;

DETHLEFS, B ;

LEE, CL ;

HALL, JE ;

GUTMAN, GA ;

CHANDY, KG .

NEURON, 1995, 15 (05) :1169-1181

[2]

ARAUJO DAM, 1993, N-S ARCH PHARMACOL, V347, P205

[3] CHARGE MOVEMENT ASSOCIATED WITH OPENING AND CLOSING OF ACTIVATION GATES OF NA CHANNELS [J].

ARMSTRONG, CM ;

BEZANILLA, F .

JOURNAL OF GENERAL PHYSIOLOGY, 1974, 63 (05) :533-552

[4] SELECTIVE BLOCKAGE OF VOLTAGE-DEPENDENT K+ CHANNELS BY A NOVEL SCORPION TOXIN [J].

CARBONE, E ;

WANKE, E ;

PRESTIPINO, G ;

POSSANI, LD ;

MAELICKE, A .

NATURE, 1982, 296 (5852) :90-91

[5] Disulfide bridge reorganization induced by proline mutations in maurotoxin [J].

Carlier, E ;

Fajloun, Z ;

Mansuelle, P ;

Fathallah, M ;

Mosbah, A ;

Oughideni, R ;

Sandoz, G ;

Di Luccio, E ;

Geib, S ;

Regaya, I ;

Brocard, J ;

Rochat, H ;

Darbon, H ;

Devaux, C ;

Sabatier, JM ;

de Waard, M .

FEBS LETTERS, 2001, 489 (2-3) :202-207

[6] PURIFICATION AND AMINO-ACID-SEQUENCES OF 6 TX3-TYPE NEUROTOXINS FROM THE VENOM OF THE BRAZILIAN ARMED SPIDER PHONEUTRIA-NIGRIVENTER (KEYS) [J].

CORDEIRO, MD ;

DEFIGUEIREDO, SG ;

VALENTIM, AD ;

DINIZ, CR ;

VONEICKSTEDT, VRD ;

GILROY, J ;

RICHARDSON, M .

TOXICON, 1993, 31 (01) :35-42

[7]

DINIZ CARLOS R., 1963, AN ACAD BRASIL CIENC, V35, P283

[8] Sea anemone peptides with a specific blocking activity against the fast inactivating potassium channel Kv3.4 [J].

Diochot, S ;

Schweitz, H ;

Béress, L ;

Lazdunski, M .

JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (12) :6744-6749

[9] Structure and pharmacology of spider venom neurotoxins [J].

Escoubas, P ;

Diochot, S ;

Corzo, G .

BIOCHIMIE, 2000, 82 (9-10) :893-907

[10] PURIFICATION AND AMINO-ACID-SEQUENCE OF THE INSECTICIDAL NEUROTOXIN TX4(6-1) FROM THE VENOM OF THE ARMED SPIDER PHONEUTRIA NIGRIVENTER (KEYS) [J].

FIGUEIREDO, SG ;

GARCIA, MELP ;

VALENTIM, AD ;

CORDEIRO, MN ;

DINIZ, CR ;

RICHARDSON, M .

TOXICON, 1995, 33 (01) :83-93

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