Cloning, expression and characterisation of an HtrA-like serine protease produced in vivo by Mycobacterium leprae.
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 degrees 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.
Laboratório de Microbiologia Celular, Instituto Oswaldo Cruz-Fiocruz, Rio de Janeiro, Brasil.
Michelle Lopes
ML
Ribeiro-Guimarães
Eliana Blini
EB
Marengo
Antonio Jorge
AJ
Tempone
Julio Jablonski
JJ
Amaral
Clécio F
CF
Klitzke
Erika K Xavier da
EK
Silveira
Fernanda Calheta Vieira
FC
Portaro
Maria Cristina Vidal
MC
Pessolani
D001483:Base Sequence; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D009166:Mycobacterium leprae / Q000201:enzymology* / Q000235:genetics; D012697:Serine Endopeptidases / Q000096:biosynthesis*; D017550:Spectroscopy, Fourier Transform Infrared
D012697:Serine Endopeptidases