Crystal structure of the holotoxino from Shigella dysenteriae at 2.5 Å resolution

Fraser, M.E.; Chernaia, Maia M.; Kozlov, Yuri; James, Michael N.G.

doi:10.1038/nsb0194-59

Cited by 276 publications

(264 citation statements)

References 25 publications

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“…1 H NMR: δ 2.38 (s, 3H), 6.85 (br s, 2H), 11.0 (br s, 1H). 13 C NMR: δ 13.8 (q), 111.2 (s), 154.6 (s), 155.6 (s), 156.1 (s), 166.4 (s). HRMS calcd for C6H7N4O2 (MH + ), 167.0569; found, 167.0573.…”

Section: -Amino-2-methyl-6h-oxazolo[54-d]pyrimidin-7-one (3)mentioning

confidence: 99%

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Structure-Based Design and Characterization of Novel Platforms for Ricin and Shiga Toxin Inhibition

et al. 2001

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Ribosome inhibiting proteins, RIPs, are a widespread family of toxic enzymes. Ricin is a plant toxin used as a poison and biological warfare agent; shiga toxin is a homologue expressed by pathogenic strains of E. coli. There is interest in creating effective antidote inhibitors to this class of enzymes. RIPs act by binding and hydrolyzing a specific adenine base from rRNA. Previous virtual screens revealed that pterins could bind in the specificity pocket of ricin and inhibit the enzyme. In this paper we explore a range of compounds that could serve as better platforms for inhibitor design. This establishes the importance of key hydrogen bond donors and acceptors for active-site complementarity. 8-Methyl-9-oxoguanine is a soluble compound that has the best inhibitory properties of any platform tested. The X-ray structure of this complex revealed that the inhibitor binds in an unexpected way that provides insight for future design. Several inhibitors of ricin were also shown to be inhibitors of shiga toxin, suggesting this program has the potential to develop effective antidotes to an important form of food poisoning.

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Section: -Amino-2-methyl-6h-oxazolo[54-d]pyrimidin-7-one (3)mentioning

confidence: 99%

“…1 H NMR (300 MHz, D2O): δ 1.88 (s). 13 C NMR (75 MHz, DMSO-d6): δ 21.9 (q), 92.6 (s), 154.5 (s), 168.2 (s), 174.9 (s). HRMS (CI) calcd for C6H9N4O3, 185.0675; found 185.0668.…”

Section: -Amino-2-methyl-6h-oxazolo[54-d]pyrimidin-7-one (3)mentioning

confidence: 99%

Structure-Based Design and Characterization of Novel Platforms for Ricin and Shiga Toxin Inhibition

et al. 2001

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“…1A). The carboxyl terminus of B-fragment was chosen since it sticks out of the globular conformation of the protein (30,31). N-Glycosylation starts in the ER by the addition of a core oligosaccharyl group from membrane-bound dolichol to an Asn-Xaa-Ser/Thr acceptor sequence on newly synthesized proteins (32).…”

Section: Shiga Toxin B-fragment With An N-glycosylation Site and The mentioning

confidence: 99%

Retrograde Transport of KDEL-bearing B-fragment of Shiga Toxin

Johannes¹,

Tenza

Antony

et al. 1997

Journal of Biological Chemistry

187

254

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To investigate retrograde transport along the biosynthetic/secretory pathway, we have constructed a recombinant Shiga toxin B-fragment carrying an N-glycosylation site and a KDEL retrieval motif at its carboxyl terminus (B-Glyc-KDEL). After incubation with HeLa cells, B-Glyc-KDEL was progressively glycosylated in the endoplasmic reticulum (ER) and remained stably associated with this compartment. B-fragment with a nonfunctional KDEL sequence (B-Glyc-KDELGL) was glycosylated with about the same kinetics as B-Glyc-KDEL but localized at steady state to the Golgi apparatus. Morphological studies showed that B-Glyc-KDEL was delivered from the plasma membrane, via endosomes and the cisternae of the Golgi apparatus, to the ER. Moreover, the addition of a sulfation site allowed us to show that B-Glyc-KDEL on transit to the ER entered the Golgi apparatus through the trans-Golgi network. Transport of B-Glyc-KDEL to the ER was slowed down by nocodazole, indicating that microtubules are important for the retrograde pathway. Our results document the existence of a continuous pathway from the plasma membrane to the endoplasmic reticulum via the Golgi apparatus and show that a fully folded exogenous protein arriving in the endoplasmic reticulum via this pathway can undergo N-glycosylation.

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“…Two types of Stx may be produced by STEC, Stx1 and/or Stx2. The Stx types all have an AB 5 structure, in which a single A-subunit is associated with five B-subunits. The A-subunit embodies the N-glycosidase catalytic activity; it acts by removing a specific adenine base from the 28 S rRNA of the 60 S ribosomal subunit within infected cells.…”

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confidence: 99%

Structure of Shiga Toxin Type 2 (Stx2) from Escherichia coli O157:H7

Fraser

Fujinaga

Cherney

et al. 2004

Journal of Biological Chemistry

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266

280

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Several serotypes of Escherichia coli produce protein toxins closely related to Shiga toxin (Stx) from Shigella dysenteriae serotype 1. These Stx-producing E. coli cause outbreaks of hemorrhagic colitis and hemolytic uremic syndrome in humans, with the latter being more likely if the E. coli produce Stx2 than if they only produce Stx1. To investigate the differences among the Stxs, which are all AB 5 toxins, the crystal structure of Stx2 from E. coli O157:H7 was determined at 1.8-Å resolution and compared with the known structure of Stx. Our major finding was that, in contrast to Stx, the active site of the A-subunit of Stx2 is accessible in the holotoxin, and a molecule of formic acid and a water molecule mimic the binding of the adenine base of the substrate. Further, the A-subunit adopts a different orientation with respect to the B-subunits in Stx2 than in Stx, due to interactions between the carboxyl termini of the B-subunits and neighboring regions of the A-subunit. Of the three types of receptor-binding sites in the B-pentamer, one has a different conformation in Stx2 than in Stx, and the carboxyl terminus of the A-subunit binds at another. Any of these structural differences might result in different mechanisms of action of the two toxins and the development of hemolytic uremic syndrome upon exposure to Stx2.

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Crystal structure of the holotoxino from Shigella dysenteriae at 2.5 Å resolution

Cited by 276 publications

References 25 publications

Structure-Based Design and Characterization of Novel Platforms for Ricin and Shiga Toxin Inhibition

Structure-Based Design and Characterization of Novel Platforms for Ricin and Shiga Toxin Inhibition

Retrograde Transport of KDEL-bearing B-fragment of Shiga Toxin

Structure of Shiga Toxin Type 2 (Stx2) from Escherichia coli O157:H7

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