Single-domain antibodies pinpoint potential targets within Shigella invasion plasmid antigen D of the needle tip complex for inhibition of type III secretion

Barta, Michael L.; Shearer, Jonathan P.; Arizmendi, Olivia; Tremblay, Jacqueline M.; Mehzabeen, Nurjahan; Zheng, Qi; Battaile, Kevin P.; Lovell, Scott; Tzipori, Saul; Picking, William D.; Shoemaker, Charles B.; Picking, Wendy L.

doi:10.1074/jbc.m117.802231

Title: Single-domain antibodies pinpoint potential targets within Shigella invasion plasmid antigen D of the needle tip complex for inhibition of type III secretion

Journal Article · Fri Aug 25 00:00:00 EDT 2017 · Journal of Biological Chemistry

DOI:https://doi.org/10.1074/jbc.m117.802231· OSTI ID:1405026

Barta, Michael L. ^[1]; Shearer, Jonathan P. ^[2]; Arizmendi, Olivia ^[1]; Tremblay, Jacqueline M. ^[2]; Mehzabeen, Nurjahan ^[1]; Zheng, Qi ^[1]; Battaile, Kevin P. ^[3]; Lovell, Scott ^[1]; Tzipori, Saul ^[2]; Picking, William D. ^[1]; Shoemaker, Charles B. ^[2]; Picking, Wendy L. ^[1]

Univ. of Kansas, Lawrence, KS (United States)
Tufts Clinical and Translational Science Institute, North Grafton, MA (United States)
Hauptman-Woodward Medical Research Institute, Argonne, IL (United States)

Numerous Gram-negative pathogens infect eukaryotes and use the type III secretion system (T3SS) to deliver effector proteins into host cells. One important T3SS feature is an extracellular needle with an associated tip complex responsible for assembly of a pore-forming translocon in the host cell membrane. Shigella spp. cause shigellosis, also called bacillary dysentery, and invade colonic epithelial cells via the T3SS. The tip complex of Shigella flexneri contains invasion plasmid antigen D (IpaD), which initially regulates secretion and provides a physical platform for the translocon pore. The tip complex represents a promising therapeutic target for many important T3SS-containing pathogens. Here, in an effort to further elucidate its function, we created a panel of single-VH domain antibodies (VHHs) that recognize distinct epitopes within IpaD. These VHHs recognized the in situ tip complex and modulated the infectious properties of Shigella. Moreover, structural elucidation of several IpaD–VHH complexes provided critical insights into tip complex formation and function. Of note, one VHH heterodimer could reduce Shigella hemolytic activity by >80%. Our observations along with previous findings support the hypothesis that the hydrophobic translocator (IpaB in Shigella) likely binds to a region within the tip protein that is structurally conserved across all T3SS-possessing pathogens, suggesting potential therapeutic avenues for managing infections by these pathogens.

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Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE Office of Science (SC); National Institutes of Health (NIH)

OSTI ID:: 1405026

Journal Information:: Journal of Biological Chemistry, Vol. 292, Issue 40; ISSN 0021-9258

Publisher:: American Society for Biochemistry and Molecular BiologyCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 9 works

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59 BASIC BIOLOGICAL SCIENCES
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
crystal structure
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Title: Single-domain antibodies pinpoint potential targets within Shigella invasion plasmid antigen D of the needle tip complex for inhibition of type III secretion

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