Ribosyl-diphthamide: Confirmation of structure by fast atom bombardment mass spectrometry

Bodley, James W.; Upham, Roger A.; Crow, Frank W.; Tomer, Kenneth B.; Gross, Michael L.

doi:10.1016/0003-9861(84)90439-9

Cited by 21 publications

(5 citation statements)

References 10 publications

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“…As we noted previously, the A fragment contains only the 1 conserved His while the B fragment has 15 (London et al, 1986). As we speculated, this pattern may somehow reflect the catalytic function of DT, which attacks a modified His residue found on elongation factor 2 and ADP-ribosylates it on the His ring (Bodley et al, 1984). PE shows a similar pattern in its sequence with only 3 His residues in its C-terminal active fragment and none in its middle domains more C-terminal than the site of the disulfide bond between residues Cys-265 and -280, but with 12 His residues in its N-terminal half which, like the B fragment of DT, is believed to carry the receptor binding site (Allured et al, 1986;Hwang et al, 1987).…”

Section: Added In Proofmentioning

confidence: 95%

Localization of the active site of diphtheria toxin

Zhao¹,

London²

1988

Biochemistry

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Information about the location of the active site of diphtheria toxin was derived from proteolysis studies and an analysis of its sequence. It was found that a specific trypsin cleavage within whole diphtheria toxin occurs at Lys-39. Therefore, Lys-39 appears to be a surface residue. Furthermore, protection from proteolysis could be obtained upon binding of either the substrate beta-nicotinamide adenine dinucleotide (oxidized form) (NAD+) or a competing ligand, adenylyl(3'-5')uridine 3'-phosphate (ApUp). The protection by ApUp, which binds to the toxin very tightly, required only stoichiometric levels. The most likely explanation of these results is that both NAD+ binding and ApUp binding block trypsin either through a steric mechanism or through a local conformational change, suggesting Lys-39 may be near the active site. Further evidence supporting this conclusion comes from comparison of the previously determined sequences of diphtheria toxin and of Pseudomonas exotoxin A, a protein that catalyzes an identical reaction. We find a significant degree of homology between the N-terminal halves of the catalytic domains of these two proteins, which apparently represents active-site residues, and that Lys-39 is in the center of the homologous sequence. Furthermore, the location of the amino acid that is the homologue of Lys-39 within the crystal structure of Pseudomonas exotoxin A is also in agreement with a location in or near the active site. Other unusual features in the sequences of diphtheria toxin and Pseudomonas exotoxin A are also described, and on the basis of the experiments presented, a possible function for ApUp is considered.

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Section: Added In Proofmentioning

confidence: 95%

Localization of the active site of diphtheria toxin

Zhao¹,

London²

1988

Biochemistry

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“…In the cytosol, the A chain catalyzes ADP-ribosylation of elongation factor 2 (EF-2), leading to inhibition of protein synthesis and cell death (5). This toxin-catalyzed activity occurs at a unique posttranslational histidine derivative, diphthamide (5,6), found in a conserved amino acid (7) sequence in the EF-2 of all eukaryotes and archaebacteria (8).…”

Section: Diphtheria Toxin (Dt)mentioning

confidence: 99%

Expression of an Anti-CD3 Single-Chain Immunotoxin with a Truncated Diphtheria Toxin in a Mutant CHO Cell Line

Liu

Gordienko

Mathias

et al. 2000

Protein Expression and Purification

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“…FAB combined with MS/MS has been applied successfully to the determination of nucleosides and nucleotides, as well as to some modified nucleosides and nucleotides (9)(10)(11)(12)(13)(14)(15)(16)(17).…”

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confidence: 99%