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

被引:18
作者
Barta, Michael L. [1 ]
Shearer, Jonathan P. [2 ]
Arizmendi, Olivia [1 ]
Tremblay, Jacqueline M. [2 ]
Mehzabeen, Nurjahan [3 ]
Zheng, Qi [1 ]
Battaile, Kevin P. [4 ]
Lovell, Scott [3 ]
Tzipori, Saul [2 ]
Picking, William D. [1 ]
Shoemaker, Charles B. [2 ]
Picking, Wendy L. [1 ]
机构
[1] Univ Kansas, Dept Pharmaceut Chem, Lawrence, KS 66047 USA
[2] Tufts Clin & Translat Sci Inst, Dept Infect Dis & Global Hlth, North Grafton, MA 02111 USA
[3] Univ Kansas, Del Shankel Struct Biol Ctr, Prot Struct Lab, Lawrence, KS 66047 USA
[4] Hauptman Woodward Med Res Inst, IMCA CAT, Argonne, IL 60439 USA
基金
美国能源部; 美国国家卫生研究院;
关键词
SYSTEM NEEDLE; PROTEIN SECRETION; IPAD; FLEXNERI; MODEL; NANOBODIES; CHALLENGE; PATHOGENS; APPARATUS; FEATURES;
D O I
10.1074/jbc.M117.802231
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:16677 / 16687
页数:11
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