Physiological Osmotic Induction of
            <i>Leptospira interrogans</i>
            Adhesion: LigA and LigB Bind Extracellular Matrix Proteins and Fibrinogen

Choy, Henry A.; Kelley, Melissa M.; Chen, Tammy L.; Møller, Anders Hauer; Matsunaga, James; Haake, David A.

doi:10.1128/iai.01635-06

Cited by 272 publications

(342 citation statements)

References 71 publications

(98 reference statements)

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“…LigB binds to Fn, fibrinogen, laminin, elastin, proelastin, and collagen and hence probably plays a substantial role in adhesion for pathogenic Leptospira (13,14,44,45). We have shown that LigBCen2 strongly binds to the NTD and weakly binds to the GBD of Fn (14) (NTD, K D ϭ 272 nM; GBD, K D ϭ 1200 nM).…”

Section: Discussionmentioning

confidence: 99%

Fibronectin Binds to and Induces Conformational Change in a Disordered Region of Leptospiral Immunoglobulin-like Protein B

Lin¹,

Greenwood

Nicholson

et al. 2009

Journal of Biological Chemistry

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Leptospira interrogans is a pathogenic spirochete that causes disease in both humans and animals. LigB (Leptospiral immunoglobulin-like protein B) contributes to the binding of Leptospira to extracellular matrix proteins such as fibronectin (Fn), fibrinogen, laminin, and collagen. A high affinity Fn-binding region of LigB has been recently localized to LigBCen2, which contains the partial eleventh and full twelfth immunoglobulin-like repeats (LigBCen2R) and 47 amino acids of the non-repeat region (LigBCen2NR) of LigB. In this study, LigBCen2NR was shown to bind to the N-terminal domain (NTD) of Fn (K D ‫؍‬ 379 nM) by an enzyme-linked immunosorbent assay and isothermal titration calorimetry. Interestingly, this sequence was not observed to adopt secondary structure by far UV circular dichroism or by differential scanning calorimetry, in agreement with computer-based secondary structure predictions. A low partition coefficient (K av ) measured with gel permeation chromatography, a high hydrodynamic radius (R h ) measured with dynamic light scattering, and the insensitivity of the intrinsic viscosity to guanidine hydrochloride treatment all suggest that LigBCen2NR possesses an extended and disordered structure. Two-dimensional 15 N-1 H HSQC NMR spectra of intact LigBCen2 in the absence and presence of NTD are consistent with these observations, suggesting the presence of both a ␤-rich region and an unstructured region in LigBCen2 and that the latter of these selectively interacts with NTD. Upon binding to NTD, LigBCen2NR was observed by CD to adopt a ␤-strand-rich structure, suggestive of the known ␤-zipper mode of NTD binding.Leptospira interrogans is a pathogenic spirochete that causes leptospirosis throughout the world, especially in developing countries but also in regions of the United States where it has reemerged (1). Weil's syndrome, a severe form of this disease, is an acute febrile illness associated with multiorgan damage, including liver failure (jaundice), renal failure (nephritis), pulmonary hemorrhage, and meningitis (1), and has a 15% mortality rate if not treated (2). The molecular pathogenesis of leptospirosis is poorly understood, and the bacterial virulence factors involved are largely unknown. Recently, several potential Leptospira virulence factors have been described, including sphingomyelinases, serine proteases, zinc-dependent proteases, and collagenase (3); LipL32 (4); lipopolysaccharide (5); a novel factor H, laminin, and Fn-binding protein (Lsa24 or Len) (6 -8); Loa 22 (9); and Lig (Leptospiral immunoglobulin-like) proteins (10 -12).Lig proteins, including LigA, LigB, and LigC, contain multiple immunoglobulin-like repeat domains (13 in LigA, 12 in LigB and LigC) (10 -12). Interestingly, the first 630 residues, from the N terminus to the first half of the seventh immunoglobulinlike domain, are conserved between LigA and LigB, but the rest of the immunoglobulin-like domains are variable (10 -12) between the two proteins. Also, a non-immunoglobulin-like repeat region found on the C-terminal...

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Section: Discussionmentioning

confidence: 99%

Fibronectin Binds to and Induces Conformational Change in a Disordered Region of Leptospiral Immunoglobulin-like Protein B

Lin¹,

Greenwood

Nicholson

et al. 2009

Journal of Biological Chemistry

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“…Indeed, several of these proteins have been reported to be leptospiral adhesins (2,4,12,28,44,49). Furthermore, well-conserved outer membrane proteins (OMPs) are promising vaccine targets, because they would have an advantage in inducing cross-protective immunity (31).…”

mentioning

confidence: 99%

OmpL1 Is an Extracellular Matrix- and Plasminogen-Interacting Protein of Leptospira spp

et al. 2012

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ABSTRACTLeptospirosis is a zoonosis with multisystem involvement caused by pathogenic strains of the genusLeptospira. OmpL1 is an outer membrane protein ofLeptospiraspp. that is expressed during infection. In this work, we investigated novel features of this protein. We describe that OmpL1 is a novel leptospiral extracellular matrix (ECM)-binding protein and a plasminogen (PLG) receptor. The recombinant protein was expressed inEscherichia coliBL21(DE3) Star/pLysS as inclusion bodies, refolded, and purified by metal-chelating chromatography. The protein presented a typical β-strand secondary structure, as evaluated by circular dichroism spectroscopy. The recombinant protein reacted with antibodies in serum samples from convalescent leptospirosis patients with a high specificity compared to serum samples from individuals with unrelated diseases. These data strengthen the usefulness of OmpL1 as a diagnostic marker of leptospirosis. The characterization of the immunogenicity of recombinant OmpL1 in inoculated BALB/c mice showed that the protein has the capacity to elicit humoral and cellular immune responses, as denoted by high antibody titers and the proliferation of lymphocytes. We demonstrate that OmpL1 has the ability to mediate attachment to laminin and plasma fibronectin, withKD(equilibrium dissociation constant) values of 2,099.93 ± 871.03 nM and 1,239.23 ± 506.85 nM, respectively. OmpL1 is also a PLG receptor, with aKDof 368.63 ± 121.23 nM, capable of generating enzymatically active plasmin. This is the first report that shows and characterizes OmpL1 as an ECM-interacting and a PLG-binding protein ofLeptospiraspp. that may play a role in bacterial pathogenesis when expressed during infection.

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“…Accordingly, leptospires employ surface-exposed proteins to bind to a wide range of host ligands, including laminin, collagen, cellular and plasma fibronectin, elastin, proteoglycans, fibrinogen, and plasminogen (13,30,31,36,37). LipL32 binds to plasma fibronectin, collagen types I, IV, and V, laminin, and plasminogen (11)(12)(13).…”

Section: Discussionmentioning

confidence: 99%

“…Several pathogenic bacteria interact with ECM components (11,12,(25)(26)(27)(28)(29)(30)(31)(32) or other host molecules such as plasminogen (13,(33)(34)(35) to facilitate host colonization. Accordingly, leptospires employ surface-exposed proteins to bind to a wide range of host ligands, including laminin, collagen, cellular and plasma fibronectin, elastin, proteoglycans, fibrinogen, and plasminogen (13,30,31,36,37).…”

Section: Discussionmentioning

confidence: 99%

Calcium Binding to Leptospira Outer Membrane Antigen LipL32 Is Not Necessary for Its Interaction with Plasma Fibronectin, Collagen Type IV, and Plasminogen

Hauk

Barbosa

et al. 2012

Journal of Biological Chemistry

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LipL32 is the most abundant outer membrane protein from pathogenic Leptospira and has been shown to bind extracellular matrix (ECM) proteins as well as Ca 2؉ . Recent crystal structures have been obtained for the protein in the apo-and Ca 2؉ -bound forms. In this work, we produced three LipL32 mutants (D163-168A, Q67A, and S247A) and evaluated their ability to interact with Ca 2؉ and with ECM glycoproteins and human plasminogen. The D163-168A mutant modifies aspartate residues involved in Ca 2؉ binding, whereas the other two modify residues in a cavity on the other side of the protein structure. Loss of calcium binding in the D163-D168A mutant was confirmed using intrinsic tryptophan fluorescence, circular dichroism, and thermal denaturation whereas the Q67A and S247A mutants presented the same Ca 2؉ affinity as the wild-type protein. We then evaluated if Ca 2؉ binding to LipL32 would be crucial for its interaction with collagen type IV and plasma proteins fibronectin and plasminogen. Surprisingly, the wild-type protein and all three mutants, including the D163-168A variant, bound to these ECM proteins with very similar affinities, both in the presence and absence of Ca 2؉ ions. In conclusion, calcium binding to LipL32 may be important to stabilize the protein, but is not necessary to mediate interaction with host extracellular matrix proteins.

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Physiological Osmotic Induction of Leptospira interrogans Adhesion: LigA and LigB Bind Extracellular Matrix Proteins and Fibrinogen

Cited by 272 publications

References 71 publications

Fibronectin Binds to and Induces Conformational Change in a Disordered Region of Leptospiral Immunoglobulin-like Protein B

Fibronectin Binds to and Induces Conformational Change in a Disordered Region of Leptospiral Immunoglobulin-like Protein B

OmpL1 Is an Extracellular Matrix- and Plasminogen-Interacting Protein of Leptospira spp

Calcium Binding to Leptospira Outer Membrane Antigen LipL32 Is Not Necessary for Its Interaction with Plasma Fibronectin, Collagen Type IV, and Plasminogen

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