Cloning, expression and characterisation of an HtrA-like serine protease produced in vivo by Mycobacterium leprae

Memórias do Instituto Oswaldo Cruz

Michelle Lopes Ribeiro-guimarães²  Eliana Blini Marengo¹  Antonio Jorge Tempone²  Julio Jablonski Amaral²  Clécio F Klitzke¹  Erika K Xavier Da Silveira²  Fernanda Calheta Vieira Portaro¹  Maria Cristina Vidal Pessolani² 

[1] ,Instituto Oswaldo Cruz-Fiocruz Laboratório de Microbiologia Celular Rio de Janeiro RJ ,Brasil

关键词: Mycobacterium leprae;    HtrA2;    protease;    enzymatic activity;    FRET peptides;    pathogenesis;   

DOI  :  10.1590/S0074-02762009000800010

来源: SciELO

【 摘 要 】

Members of the high temperature requirement A (HtrA) family of chaperone proteases have been shown to play a role in bacterial pathogenesis. In a recent report, we demonstrated that the gene ML0176, which codes for a predicted HtrA-like protease, a gene conserved in other species of mycobacteria, is transcribed by Mycobacterium leprae in human leprosy lesions. In the present study, the recombinant ML0176 protein was produced and its enzymatic properties investigated. M. lepraerecombinant ML0176 was able to hydrolyse a variety of synthetic and natural peptides. Similar to other HtrA proteins, this enzyme displayed maximum proteolytic activity at temperatures above 40°C and was completely inactivated by aprotinin, a protease inhibitor with high selectivity for serine proteases. Finally, analysis of M. leprae ML0176 specificity suggested a broader cleavage preference than that of previously described HtrAs homologues. In summary, we have identified an HtrA-like protease in M. lepraethat may constitute a potential new target for the development of novel prophylactic and/or therapeutic strategies against mycobacterial infections.

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