title: Fibronectin binds to and induces conformational change in a disordered region of leptospiral immunoglobulin-like protein B.
abstract: 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 beta-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 beta-strand-rich structure, suggestive of the known beta-zipper mode of NTD binding.
authors: Department of Population Medicine, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
authors: Yi-Pin
authors: YP
authors: Lin
authors: Alex
authors: A
authors: Greenwood
authors: Linda K
authors: LK
authors: Nicholson
authors: Yogendra
authors: Y
authors: Sharma
authors: Sean P
authors: SP
authors: McDonough
authors: Yung-Fu
authors: YF
authors: Chang
mesh_terms: D000942:Antigens, Bacterial / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005353:Fibronectins / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D007921:Leptospira interrogans / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D007922:Leptospirosis / Q000378:metabolism* / Q000382:microbiology; D011485:Protein Binding; D017434:Protein Structure, Tertiary
chemical_list: D000942:Antigens, Bacterial; D005353:Fibronectins; C487006:LigB-m protein, Leptospira interrogans