Expression and characterization of a recombinant cysteine proteinase of Leishmania mexicana

Expression and characterization of a recombinant cysteine proteinase of Leishmania mexicana

Autor Sanderson, Sanya J. Google Scholar
Pollock, Kevin GJ Google Scholar
Hilley, James D. Google Scholar
Meldal, Morten Google Scholar
St Hilaire, Phaedria Google Scholar
Juliano, Maria Aparecida Autor UNIFESP Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Mottram, Jeremy C. Google Scholar
Coombs, Graham H. Google Scholar
Instituição Univ Glasgow
Carlsberg Lab
Universidade Federal de São Paulo (UNIFESP)
Resumo A major cysteine proteinase (CPB) of Leishmania mexicana, that is predominantly expressed in the form of the parasite that causes disease in mammals, has been overexpressed in Escherichia coli and purified from inclusion bodies to apparent homogeneity. the CPB enzyme, CPB2.8, was expressed as an inactive pro-form lacking the characteristic C-terminal extension (CPB2.8 Delta CTE). Pro-region processing was initiated during protein refolding and proceeded through several intermediate stages. Maximum enzyme activity accompanied removal of the entire pro-region. This was facilitated by acidification. Purified mature enzyme gave a single band on SDS/PAGE and gelatin SDS/PAGE gels, co-migrated with native enzyme in L. mexicana lysates, and had the same N-terminal sequence as the native enzyme. the procedure yielded > 3.5 mg of active enzyme per litre of E. coli culture.
Palavra-chave cathepsin L
pro-region processing
recombinant enzyme
Idioma Inglês
Data de publicação 2000-04-15
Publicado em Biochemical Journal. London: Portland Press, v. 347, p. 383-388, 2000.
ISSN 0264-6021 (Sherpa/Romeo, fator de impacto)
Publicador Portland Press
Extensão 383-388
Fonte http://dx.doi.org/10.1042/0264-6021:3470383
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:000086792800008
Endereço permanente http://repositorio.unifesp.br/handle/11600/26295

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