Repeated domains of leptospira immunoglobulin-like proteins interact with elastin and tropoelastin.
Leptospira spp., the causative agents of leptospirosis, adhere to components of the extracellular matrix, a pivotal role for colonization of host tissues during infection. Previously, we and others have shown that Leptospira immunoglobulin-like proteins (Lig) of Leptospira spp. bind to fibronectin, laminin, collagen, and fibrinogen. In this study, we report that Leptospira can be immobilized by human tropoelastin (HTE) or elastin from different tissues, including lung, skin, and blood vessels, and that Lig proteins can bind to HTE or elastin. Moreover, both elastin and HTE bind to the same LigB immunoglobulin-like domains, including LigBCon4, LigBCen7'-8, LigBCen9, and LigBCen12 as demonstrated by enzyme-linked immunosorbent assay (ELISA) and competition ELISAs. The LigB immunoglobulin-like domain binds to the 17th to 27th exons of HTE (17-27HTE) as determined by ELISA (LigBCon4, K(D) = 0.50 microm; LigBCen7'-8, K(D) = 0.82 microm; LigBCen9, K(D) = 1.54 microm; and LigBCen12, K(D) = 0.73 microm). The interaction of LigBCon4 and 17-27HTE was further confirmed by steady state fluorescence spectroscopy (K(D) = 0.49 microm) and ITC (K(D) = 0.54 microm). Furthermore, the binding was enthalpy-driven and affected by environmental pH, indicating it is a charge-charge interaction. The binding affinity of LigBCon4D341N to 17-27HTE was 4.6-fold less than that of wild type LigBCon4. In summary, we show that Lig proteins of Leptospira spp. interact with elastin and HTE, and we conclude this interaction may contribute to Leptospira adhesion to host tissues during infection.
Lin
Yi-Pin
YP
Department of Population Medicine and Diagnostic Sciences, Cornell University, Ithaca, New York, USA.
Lee
Dae-Won
DW
McDonough
Sean P
SP
Nicholson
Linda K
LK
Sharma
Yogendra
Y
Chang
Yung-Fu
YF
D000595:Amino Acid Sequence; D001011:Aorta / Q000737:chemistry; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites / Q000235:genetics; D019921:Biotinylation; D018914:Cells, Immobilized; D004549:Elastin / Q000235:genetics / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics / Q000378:metabolism; D007921:Leptospira interrogans / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D008168:Lung / Q000737:chemistry; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D012867:Skin / Q000737:chemistry; D014333:Tropoelastin / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D007136:Immunoglobulins; D014333:Tropoelastin; D004549:Elastin