Mass Fingerprinting of the Venom and Transcriptome of Venom Gland of Scorpion Centruroides tecomanus

被引:54
作者
Valdez-Velazquez, Laura L. [1 ,2 ]
Quintero-Hernandez, Veronica [3 ]
Teresa Romero-Gutierrez, Maria [1 ,2 ]
Coronas, Fredy I. V. [3 ]
Possani, Lourival D. [3 ]
机构
[1] Univ Colima, Fac Ciencias Quim, Colima, Mexico
[2] Univ Colima, Fac Med, Colima, Mexico
[3] Univ Nacl Autonoma Mexico, Inst Biotecnol, Cuernavaca 62191, Morelos, Mexico
关键词
AMINO-ACID-SEQUENCE; TITYUS-SERRULATUS SCORPIONES; RICH ANTIMICROBIAL PEPTIDES; MEXICAN SCORPION; SPECTROMETRY ANALYSIS; MOLECULAR DIVERSITY; PROTEOMIC ANALYSIS; BETA-TOXINS; CHANNELS; PURIFICATION;
D O I
10.1371/journal.pone.0066486
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Centruroides tecomanus is a Mexican scorpion endemic of the State of Colima, that causes human fatalities. This communication describes a proteome analysis obtained from milked venom and a transcriptome analysis from a cDNA library constructed from two pairs of venom glands of this scorpion. High perfomance liquid chromatography separation of soluble venom produced 80 fractions, from which at least 104 individual components were identified by mass spectrometry analysis, showing to contain molecular masses from 259 to 44,392 Da. Most of these components are within the expected molecular masses for Na+- and K+-channel specific toxic peptides, supporting the clinical findings of intoxication, when humans are stung by this scorpion. From the cDNA library 162 clones were randomly chosen, from which 130 sequences of good quality were identified and were clustered in 28 contigs containing, each, two or more expressed sequence tags (EST) and 49 singlets with only one EST. Deduced amino acid sequence analysis from 53% of the total ESTs showed that 81% (24 sequences) are similar to known toxic peptides that affect Na+-channel activity, and 19% (7 unique sequences) are similar to K+-channel especific toxins. Out of the 31 sequences, at least 8 peptides were confirmed by direct Edman degradation, using components isolated directly from the venom. The remaining 19%, 4%, 4%, 15% and 5% of the ESTs correspond respectively to proteins involved in cellular processes, antimicrobial peptides, venom components, proteins without defined function and sequences without similarity in databases. Among the cloned genes are those similar to metalloproteinases.
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页数:15
相关论文
共 72 条
[1]   Profiling the resting venom gland of the scorpion Tityus stigmurus through a transcriptomic survey [J].
Almeida, Diego D. ;
Scortecci, Katia C. ;
Kobashi, Leonardo S. ;
Agnez-Lima, Lucymara F. ;
Medeiros, Silvia R. B. ;
Silva-Junior, Arnobio A. ;
Junqueira-de-Azevedo, Inacio de L. M. ;
Fernandes-Pedrosa, Matheus de F. .
BMC GENOMICS, 2012, 13
[2]   Enzymes with gelatinolytic activity can be found in Tityus bahiensis and Tityus serrulatus venoms [J].
Almeida, FM ;
Pimenta, AMC ;
De Figueiredo, SG ;
Santoro, MM ;
Martin-Eauclaire, MF ;
Diniz, CR ;
De Lima, ME .
TOXICON, 2002, 40 (07) :1041-1045
[3]  
[Anonymous], 2000, CATALOG SCORPIONS WO
[4]   Proteomic analysis of the venom from the scorpion Tityus stigmurus:: Biochemical and physiological comparison with other Tityus species [J].
Batista, C. V. F. ;
Roman-Gonzalez, S. A. ;
Salas-Castillo, S. P. ;
Zamudio, F. Z. ;
Gomez-Lagunas, F. ;
Possani, L. D. .
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY, 2007, 146 (1-2) :147-157
[5]   Proteomics of the venom from the Amazonian scorpion Tityus cambridgei and the role of prolines on mass spectrometry analysis of toxins [J].
Batista, CVF ;
del Pozo, L ;
Zamudio, FZ ;
Contreras, S ;
Becerril, B ;
Wanke, E ;
Possani, LD .
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2004, 803 (01) :55-66
[6]   CLONING AND CHARACTERIZATION OF CDNAS THAT CODE FOR NA+-CHANNEL-BLOCKING TOXINS OF THE SCORPION CENTRUROIDES-NOXIUS HOFFMANN [J].
BECERRIL, B ;
VAZQUEZ, A ;
GARCIA, CI ;
CORONA, M ;
BOLIVAR, F ;
POSSANI, LD .
GENE, 1993, 128 (02) :165-171
[7]   Recombinant scorpine:: a multifunctional antimicrobial peptide with activity against different pathogens [J].
Carballar-Lejarazu, R. ;
Rodriguez, M. H. ;
Hernandez-Hernandez, F. de la Cruz ;
Ramos-Castaneda, J. ;
Possani, L. D. ;
Zurita-Ortega, M. ;
Reynaud-Garza, E. ;
Hernandez-Rivas, R. ;
Loukeris, T. ;
Lycett, G. ;
Lanz-Mendoza, H. .
CELLULAR AND MOLECULAR LIFE SCIENCES, 2008, 65 (19) :3081-3092
[8]   Voltage-gated ion channels and gating modifier toxins [J].
Catterall, William A. ;
Cestele, Sandrine ;
Yarov-Yarovoy, Vladimir ;
Yu, Frank H. ;
Konoki, Keiichi ;
Scheuer, Todd .
TOXICON, 2007, 49 (02) :124-141
[9]   ALPHA-SCORPION TOXINS BINDING ON RAT-BRAIN AND INSECT SODIUM-CHANNELS REVEAL DIVERGENT ALLOSTERIC MODULATIONS BY BREVETOXIN AND VERATRIDINE [J].
CESTELE, S ;
KHALIFA, RB ;
PELHATE, M ;
ROCHAT, H ;
GORDON, D .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (25) :15153-15161
[10]   Epidemiology of scorpionism: A global appraisal [J].
Chippaux, J. -P. ;
Goyffon, M. .
ACTA TROPICA, 2008, 107 (02) :71-79