Crystal Structure of LipL32, the Most Abundant Surface Protein of Pathogenic Leptospira spp.

Vivian, J.P.; Beddoe, Travis; McAlister, Adrian Dale; Wilce, Matthew Charles James; Zaker-Tabrizi, Leyla; Troy, S.; Byres, E.; Hoke, David E.; Cullen, Paul; Lo, Miranda; Murray, Gerald L; Adler, Ben; Rossjohn, Jamie

doi:10.1016/j.jmb.2009.02.038

Cited by 52 publications

(46 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These PTM sites are localized in the regions containing two electronegative patches (dominated by Glu 250 , Glu 251 , Glu 260 and Glu 261 , and by Glu 138 , Asp 167 and Asp 168 ) revealed by the crystal structure of LipL32 determined in the recent study [39]. These two acid-rich patches were identified as potential binding sites for extracellular matrix (ECM) proteins such as laminin.…”

Section: Biological Relevance Of Modified Proteins In L Interrogansmentioning

confidence: 97%

High-coverage proteome analysis reveals the first insight of protein modification systems in the pathogenic spirochete Leptospira interrogans

Cao

Dai

et al. 2009

Cell Res

View full text Add to dashboard Cite

show abstract

Section: Biological Relevance Of Modified Proteins In L Interrogansmentioning

confidence: 97%

High-coverage proteome analysis reveals the first insight of protein modification systems in the pathogenic spirochete Leptospira interrogans

Cao

Dai

et al. 2009

Cell Res

View full text Add to dashboard Cite

show abstract

“…LipL32 is expressed in all pathogenic Leptospira spp., and it is highly conserved (19) and not expressed in the saprophytic L. biflexa (29). This protein binds to extracellular matrix components, as indicated by in vitro assays (22,23) and crystal structure analyses (36). Moreover, LipL32 is expressed during mammalian leptospiral infection (18).…”

mentioning

confidence: 99%

Protection against Lethal Leptospirosis after Vaccination with LipL32 Coupled or Coadministered with the B Subunit of Escherichia coli Heat-Labile Enterotoxin

Grassmann

Félix

Santos

et al. 2012

Clin. Vaccine Immunol.

View full text Add to dashboard Cite

ABSTRACTLeptospirosis, a worldwide zoonosis, lacks an effective, safe, and cross-protective vaccine. LipL32, the most abundant, immunogenic, and conserved surface lipoprotein present in all pathogenic species ofLeptospira, is a promising antigen candidate for a recombinant vaccine. However, several studies have reported a lack of protection when this protein is used as a subunit vaccine. In an attempt to enhance the immune response, we used LipL32 coupled to or coadministered with the B subunit of theEscherichia coliheat-labile enterotoxin (LTB) in a hamster model of leptospirosis. After homologous challenge with 5× the 50% lethal dose (LD50) ofLeptospira interrogans, animals vaccinated with LipL32 coadministered with LTB and LTB::LipL32 had significantly higher survival rates (P< 0.05) than animals from the control group. This is the first report of a protective immune response afforded by a subunit vaccine using LipL32 and represents an important contribution toward the development of improved leptospirosis vaccines.

show abstract

“…Recently, Vieira et al (13) showed that LipL32 is able to bind to human plasminogen, the inactive precursor of the extracellular proteinase plasmin. To better understand the biological role and structural requirements for the function of this important lipoprotein, the crystal structure of recombinant LipL32 was initially determined in the apo state (14,15). Our previous observation that many of the proteins with the closest structural homology to LipL32 bind both calcium ions and extracellular matrix proteins led us to demonstrate for the first time that LipL32 binds Ca 2ϩ , but not other divalent metals such as Mg 2ϩ , Zn 2ϩ , and Cu 2ϩ (14).…”

Section: Discussionmentioning

confidence: 99%

“…Mutants-The resolution of the apoLipL32 structure (14,15) led to the prediction and demonstration of its ability to bind Ca 2ϩ (14). Because the apo-structure did not unambiguously reveal the Ca 2ϩ -binding site, we produced three LipL32 mutants designed to disrupt two originally proposed candidate binding sites on opposite sides of the protein: (i) LipL32 Q67A and LipL32 S247A have mutations in a cavity that coincides with the Ca 2ϩ -binding site from the ColG collagenase from Clostridium histolyticum and (ii) LipL32 D163-168A has five aspartates in a conserved acidic loop all changed to alanines.…”

Section: Production Of Lipl32mentioning

confidence: 99%

“…The crystal structure of apoLipL32 (14,15) and the calcium-bound structure (16) led to the characterization of the significant structural changes associated with metal binding that includes strand-coil transitions and a 10-Å translocation of an aspartate-rich loop to form part of the Ca 2ϩ -binding site. These observations raised the question regarding the role of Ca 2ϩ in the binding of LipL32 to its target proteins.…”

mentioning

confidence: 99%

See 1 more Smart Citation

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

View full text Add to dashboard Cite

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.

show abstract

Crystal Structure of LipL32, the Most Abundant Surface Protein of Pathogenic Leptospira spp.

Cited by 52 publications

References 45 publications

High-coverage proteome analysis reveals the first insight of protein modification systems in the pathogenic spirochete Leptospira interrogans

High-coverage proteome analysis reveals the first insight of protein modification systems in the pathogenic spirochete Leptospira interrogans

Protection against Lethal Leptospirosis after Vaccination with LipL32 Coupled or Coadministered with the B Subunit of Escherichia coli Heat-Labile Enterotoxin

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

Contact Info

Product

Resources

About