A lethal cardiotoxic-cytotoxic protein from the Indian monocellate cobra (Naja kaouthia) venom

被引：52

作者：

Debnath, Anindita ^{[1
]}

Saha, Archita ^{[1
]}

Gomes, Antony ^{[2
]}

Biswas, Sumit ^{[3
]}

Chakrabarti, Pinakpani ^{[3
]}

Giri, Biplab ^{[2
]}

Biswas, Ajoy K. ^{[2
]}

Das Gupta, Shubho ^{[1
]}

Gomes, Aparna ^{[1
]}

机构：

[1] Indian Inst Chem Biol, Drug Dev Diagnost & Biotechnol Div, Kolkata 700032, India

[2] Univ Calcutta, Dept Physiol, Lab Toxinol & Expt Pharmacodynam, Kolkata 700009, India

[3] Bose Inst, Dept Biochem, Kolkata 700054, India

来源：

TOXICON | 2010年 / 56卷 / 04期

关键词：

Snake venom; Naja kaouthia; Cardiotoxin; Cytotoxin; Apoptosis; FLOW-CYTOMETRIC DETECTION; AMINO-ACID-SEQUENCE; PHOSPHATIDYLSERINE EXPRESSION; CYTOCHROME-C; SNAKE-VENOM; ANNEXIN-V; CELLS; APOPTOSIS; PROCASPASE-9; INDUCTION;

D O I：

10.1016/j.toxicon.2010.05.016

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

A lethal cardiotoxic-cytotoxic protein (mol wt. 6.76 kDa) has been purified from the Indian monocellate cobra (Naja kaouthia) venom by ion-exchange chromatography and HPLC. CD spectra indicated the presence of 23% alpha helix, 19% beta sheets and 35% coil. Complete amino acid sequence was determined by MALDI, which showed similar homology with cardiotoxins/cytotoxins isolated from venom of other Naja species. Intraperitoneal LD50 was 2 5 mg kg(-1) in BalbC male mice. In vitro cardiotoxicity studies on isolated guinea pig auricle showed that the molecule produced auricular blockade that was abolished after trypsin treatment. Cytotoxicity studies on human leukemic U937 and K562 cells showed that it significantly inhibited cell proliferation in a dose and time dependent manner, as observed by trypan blue exclusion method and tetrazolium bromide reduction assay IC50 on U937 and K562 cells were 3.5 mu g/ml and 11 mu g/ml respectively Morphometry and cell sorting studies indicated apoptosis induction in toxin treated leukemic cells Apoptosis was caspase 3 and 9 dependent and the treated leukemic cells were arrested in sub-G1 stage There was an increase in Bax-Bcl2 ratio, decrease in HSP (Heat shock protein) 70 and HSP90 and induction of PARP cleavage after NK-CTI treatment. The toxin showed low cytotoxic effect on normal human leukocytes as compared with imatinib mesylate. Further detailed cytotoxic and cardiotoxic effects at the molecular level are in progress. (C) 2010 Elsevier Ltd. All rights reserved

引用

页码：569 / 579

页数：11

共 31 条

[1] Three-dimensional structure of the apoptosome: Implications for assembly, procaspase-9 binding, and activation [J].

Acehan, D ;

Jiang, XJ ;

Morgan, DG ;

Heuser, JE ;

Wang, XD ;

Akey, CW .

MOLECULAR CELL, 2002, 9 (02) :423-432

[2] INHIBITION OF HUMAN IMMUNODEFICIENCY VIRUS IN EARLY INFECTED AND CHRONICALLY INFECTED-CELLS BY ANTISENSE OLIGODEOXYNUCLEOTIDES AND THEIR PHOSPHOROTHIOATE ANALOGS [J].

AGRAWAL, S ;

IKEUCHI, T ;

SUN, D ;

SARIN, PS ;

KONOPKA, A ;

MAIZEL, J ;

ZAMECNIK, PC .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (20) :7790-7794

[3] Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome [J].

Beere, HM ;

Wolf, BB ;

Cain, K ;

Mosser, DD ;

Mahboubi, A ;

Kuwana, T ;

Tailor, P ;

Morimoto, RI ;

Cohen, GM ;

Green, DR .

NATURE CELL BIOLOGY, 2000, 2 (08) :469-475

[4] Role of poly(ADP-ribose) polymerase (PARP) cleavage in apoptosis - Caspase 3-resistant PARP mutant increases rates of apoptosis in transfected cells [J].

Boulares, AH ;

Yakovlev, AG ;

Ivanova, V ;

Stoica, BA ;

Wang, GP ;

Iyer, S ;

Smulson, M .

JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (33) :22932-22940

[5]

BURN JH, 1952, PRACTICAL PHARM, V22, P30

[6]

Calmettr A, 1933, CR HEBD ACAD SCI, V197, P205

[7] Chemical induction of cellular antioxidants affords marked protection against oxidative injury in vascular smooth muscle cells [J].

Cao, ZX ;

Li, YB .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2002, 292 (01) :50-57

[8] Venom of Indian monocellate cobra and Russell's viper show anticancer activity in experimental models [J].

Debnath, Anindita ;

Chatterjee, Uttora ;

Das, Minati ;

Vedasiromoni, Joseph R. ;

Gomes, Aparna .

JOURNAL OF ETHNOPHARMACOLOGY, 2007, 111 (03) :681-684

[9] STRUCTURE AND PHARMACOLOGY OF ELAPID CYTOTOXINS [J].

DUFTON, MJ ;

HIDER, RC .

PHARMACOLOGY & THERAPEUTICS, 1988, 36 (01) :1-&

[10] Cancer cell injury by cytotoxins from cobra venom is mediated through lysosomal damage [J].

Feofanov, AV ;

Sharonov, GV ;

Astapova, MV ;

Rodionov, DI ;

Utkin, YN ;

Arseniev, AS .

BIOCHEMICAL JOURNAL, 2005, 390 (390) :11-18

← 1 2 3 4 →