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Journal of the Mexican Chemical Society

Print version ISSN 1870-249X

J. Mex. Chem. Soc vol.49 n.2 México  2005

 

Article

 

Amino Acid Sequence Determination and Chemical Synthesis of CllErgl (y-KTx1.5), a K+ Channel Blocker Peptide Isolated from the Scorpion Centruroides limpidus limpidus

 

Fredy I. Coronas, Cipriano Balderas, Liliana Pardo-López, Lourival D. Possani and Georgina B. Gurrola*

 

Department of Molecular Medicine and Bioprocesses, Institute of Biotechnology, National Autonomous University of México, Avenida Universidad, 2001,Cuernavaca 62210, Mexico. * e-mail: georgina@ibt.unam.mx

 

Received: November 9, 2004
Published on the web: April 12, 2005

 

Abstract

A novel toxin named CllErgl (systematic nomenclature y-KTxl.5) was purified from the venom of the scorpion Centruroides limpidus limpidus and its amino acid sequence was determined. It has 42 amino-acid residues cross-linked by four disulfide bridges and blocks specifically a potassium channel of the family ether-a-go-go (ERG). The full peptide was chemically synthesized and properly folded, showing that it blocks the human ERG-channels (HERG) with identical affinity to that of the native peptide. Synthetic CllErg1 can be produced in quantities enough to compensate its low concentration in the natural venom. It paves the way to conduct studies aimed at the identification of the structural motifs of HERG critical for proper channel function. Additionally, another analogous peptide CllErg2 (systematic name y-KTx4.1) was purified and had its full amino acid sequence determined. It contained 43 amino acid residues, maintained closely packed by four disulfide bridges.

Keywords: ERG, Centruroides limpidus limpidus, chemical synthesis, K+-channel, scorpion toxin

 

Resumo

Uma nova toxina denominada CllErgl (nomenclatura sistemática y-KTx1.5) foi purificada do veneno do escorpião Centruroides limpidus limpidus e a sua seqüência de amino ácidos foi determinada. Ela tem 42 resíduos de amino ácidos entrecruzados por quatro pontes de disulfetos e bloqueia especificamente um canal de potássio da família eter-a-go-go (ERG). O peptídeo completo foi quimicamente sintetizado e adequadamente enrolado, mostrando que é capaz de bloquear o canal-ERG humano (HERG) com uma afinidade idêntica ao peptídeo nativo. А toxina CllErgl sintética pode ser produzida em quantidades suficientes para compensar a sua baixa concentração encontrada no veneno natural. Isto prepara o caminho para conduzir estudos enfocando a identificação dos padrões estruturais do HERG que são críticos para o funcionamento adequado do canal iònico. Adicionalmente, outro peptídeo análogo CllErg2 (nome sistemático y-KTx4.1) foi purificado e a sua seqüência completa de amino ácidos foi determinada. Este contém 4З resíduos de amino ácidos, mantidos de forma compacta por quatro pontes de disulfetos.

 

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Acknowledgements

The technical support received from Dr. Fernando Zamudio, during sequencing is greatly appreciated. Thauthors are also indebted to Ing. Arturo Ocadiz Ramírez and B.Sc. Alma L. Martínez Valle for their technical support at the Computer Center of our Institute. This work was partially supported by grant number 40251 Q from the National Council of Science and Technology (CONACyT), Mexican Government and grant number IN206003 from Dirección General de Asuntos del Personal Académico of the National Autonomous University of Mexico given to LDP and GBG.

 

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