LipL53, a temperature regulated protein from Leptospira interrogans that binds to extracellular matrix molecules.
The regulation of gene expression by environmental signals, such as temperature and osmolarity, has been correlated with virulence. In this study, we characterize the protein LipL53 from Leptospira interrogans, previously shown to react with serum sample of individual diagnosed with leptospirosis and to be up-regulated by shift to physiological osmolarity. The recombinant protein was expressed in Escherichia coli system, in insoluble form, recovered by urea solubilization and further refolded by decreasing the denaturing agent concentration during the purification procedure. The secondary structure content of the recombinant LipL53, as assessed by circular dichroism, showed a mixture of beta-strands and alpha-helix. The presence of LipL53 transcript at 28 degrees C was only detected within the virulent strains. However, upon shifted of attenuated cultures of pathogenic strains from 28 degrees C to 37 degrees C and to 39 degrees C, this transcript could also be observed. LipL53 binds laminin, collagen IV, cellular and plasma fibronectin in dose-dependent and saturable manner. Animal challenge studies showed that LipL53, although immunogenic, elicited only partial protection in hamsters. LipL53 is probably surface exposed as seen through immunofluorescence confocal microscopy. Our results suggest that LipL53 is a novel temperature regulated adhesin of L. interrogans that may be relevant in the leptospiral pathogenesis.
Oliveira
Tatiane R
TR
Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil.
Longhi
Mariana T
MT
Gonçales
Amane P
AP
de Morais
Zenaide M
ZM
Vasconcellos
Silvio A
SA
Nascimento
Ana L T O
AL
D018829:Adhesins, Bacterial / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D002942:Circular Dichroism; D006224:Cricetinae; D004926:Escherichia coli / Q000235:genetics; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D007921:Leptospira interrogans / Q000472:pathogenicity*; D007922:Leptospirosis / Q000276:immunology / Q000517:prevention & control; D008297:Male; D008647:Mesocricetus; D051379:Mice; D008807:Mice, Inbred BALB C; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D013696:Temperature; D037521:Virulence Factors / Q000378:metabolism*
D018829:Adhesins, Bacterial; D011994:Recombinant Proteins; D037521:Virulence Factors