6YHK

Crystal structure of full-length CNFy (C866S) from Yersinia pseudotuberculosis

Summary for 6YHK

• Entry DOI: 10.2210/pdb6yhk/pdb
• Descriptor: Cytotoxic necrotizing factor, SULFATE ION, CHLORIDE ION, ... (4 entities in total)
• Functional Keywords: toxin, cnf, cytotoxic necrotizing factor, deamidase, rhoa modification, rhoa activation, putative adp-ribosyltransferase
• Biological source: Yersinia pseudotuberculosis
• Total number of polymer chains: 1
• Total formula weight: 115820.25

Authors

Lukat, P.,Gazdag, E.M.,Heidler, T.V.,Blankenfeldt, W. (deposition date: 2020-03-30, release date: 2020-12-30, Last modification date: 2024-05-01)

Primary citation

Chaoprasid, P.,Lukat, P.,Muhlen, S.,Heidler, T.,Gazdag, E.M.,Dong, S.,Bi, W.,Ruter, C.,Kirchenwitz, M.,Steffen, A.,Jansch, L.,Stradal, T.E.B.,Dersch, P.,Blankenfeldt, W.
Crystal structure of bacterial cytotoxic necrotizing factor CNF Y reveals molecular building blocks for intoxication.
Embo J., 40:e105202-e105202, 2021

PubMed Abstract: Cytotoxic necrotizing factors (CNFs) are bacterial single-chain exotoxins that modulate cytokinetic/oncogenic and inflammatory processes through activation of host cell Rho GTPases. To achieve this, they are secreted, bind surface receptors to induce endocytosis and translocate a catalytic unit into the cytosol to intoxicate host cells. A three-dimensional structure that provides insight into the underlying mechanisms is still lacking. Here, we determined the crystal structure of full-length Yersinia pseudotuberculosis CNF . CNF consists of five domains (D1-D5), and by integrating structural and functional data, we demonstrate that D1-3 act as export and translocation module for the catalytic unit (D4-5) and for a fused β-lactamase reporter protein. We further found that D4, which possesses structural similarity to ADP-ribosyl transferases, but had no equivalent catalytic activity, changed its position to interact extensively with D5 in the crystal structure of the free D4-5 fragment. This liberates D5 from a semi-blocked conformation in full-length CNF , leading to higher deamidation activity. Finally, we identify CNF translocation modules in several uncharacterized fusion proteins, which suggests their usability as a broad-specificity protein delivery tool.

PubMed: 33410511
DOI: 10.15252/embj.2020105202

Experimental method

X-RAY DIFFRACTION (2.7 Å)