Structure-Function Analysis of the Diphtheria Toxin Receptor Toxin Binding Site by Site-directed Mutagenesis

Mitamura, Teruaki; Umata, Toshiyuki; Nakano, Fumie; Shishido, Yuji; Toyoda, Tomohiko; Itai, Akiko; Kimurâ, Hiroshi; Mekada, Eisuke

doi:10.1074/jbc.272.43.27084

Cited by 47 publications

(36 citation statements)

References 41 publications

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“…6D), the A-loop and C-loop face the receptor-binding domain of DT (44). We previously demonstrated that the amino acids Phe 115 , Leu 127 , or especially Glu 141 , within the EGF-like domain are critical for DT-binding activity (38). In particular, Phe 115 , which is located in the A-loop, and Glu 141 , which is within the C-loop, play crucial roles in binding between DT and HB-EGF at the interface between these molecules (44).…”

Section: Discussionmentioning

confidence: 99%

“…Specific binding was determined by subtracting the nonspecific binding from the total binding obtained using 125 I-DT alone. The relative values of 125 I-DT binding to pro-HB-EGF and to pro-⌬HB on the cell surface were calculated as the ratio of specific DT binding to the total amount of pro-HB-EGF or pro-⌬HB on the cell surface, measured by the anti-HB-EGF antibody (H-6) binding assay, as previously described (38).…”

Section: Methodsmentioning

confidence: 99%

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Suppression of the Biological Activities of the Epidermal Growth Factor (EGF)-like Domain by the Heparin-binding Domain of Heparin-binding EGF-like Growth Factor

Takazaki

Shishido²,

Iwamoto

et al. 2004

Journal of Biological Chemistry

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Heparin-binding EGF-like growth factor (HB-EGF) is a member of the EGF family of growth factors that has a high affinity for heparin and heparan sulfate. While interactions with heparin are thought to modulate the biological activity of HB-EGF, the precise role of the heparin-binding domain has remained unclear. We analyzed the activity of wild-type HB-EGF and a mutant form lacking the heparin-binding domain (⌬HB) in the presence or absence of heparin. The activity of the EGFlike domain of HB-EGF was determined by measuring binding to diphtheria toxin (DT) as well as the growth factor activity in EGF receptor-expressing cells. The binding affinity of ⌬HB for DT was much higher than that of wild-type HB-EGF in the absence of heparin. The binding affinity of HB-EGF for DT was increased by addition of exogenous heparin and reached the level close to the affinity of ⌬HB, whereas that of ⌬HB was not affected. Moreover, the growth factor activity of ⌬HB was much higher than that of wild-type HB-EGF in the absence of heparin but was not affected by addition of exogenous heparin, whereas HB-EGF had increased growth factor activity with added heparin. These results indicate that the heparin-binding domain suppresses the activity of the EGF-like domain of HB-EGF and that association of heparin with HB-EGF via this domain removes the suppressive effect. Thus, we conclude that the heparin-binding domain serves as a negative regulator of this growth factor.Cell surface heparan sulfate proteoglycans (HSPGs) 1 have been implicated in a variety of cell signaling pathways involving heparin-binding growth factors or cytokines. These growth factors and cytokines form tight complexes with heparin and

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Suppression of the Biological Activities of the Epidermal Growth Factor (EGF)-like Domain by the Heparin-binding Domain of Heparin-binding EGF-like Growth Factor

Takazaki

Shishido²,

Iwamoto

et al. 2004

Journal of Biological Chemistry

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“…Binding of CRM197, the nontoxic and catalytically inactive [Glu-52] mutant of diphtheria toxin, to the extracellular HB-EGF domain potently and specifically inhibits the mitogenic activity of HB-EGF (26). Unlike in mice and rats, the amino acid residues that are critical for diphtheria toxin binding, Phe-115, Leu-127, and Glu-141 in the binding domain of the porcine proHB-EGF, are identical to those of human proHB-EGF, enabling CRM197 to bind and interact with porcine proHB-EGF (27). To determine whether HB-EGF shedding contributed to Ang II-mediated activation of EGFR, we assessed the effect of CRM197 on Ang II-mediated EGFR tyrosine phosphorylation.…”

Section: Release Of Soluble Hb-egf Mediates Ang Ii-induced Egfr Activmentioning

confidence: 99%

Role of EGF Receptor Activation in Angiotensin II–Induced Renal Epithelial Cell Hypertrophy

Chen¹,

Chen²,

Neilson³

et al. 2006

Journal of the American Society of Nephrology

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“…Diphtheria toxin is a single polypeptide of 535 amino acids and a molecular mass of 58.4 kDa, divided into two subunitssubunit A (catalytic domain) and subunit B (binding domain)-which are linked by a disulfide bond [81]. Because of this two-component structure it is structurally classified as an A-B type toxin.…”

Section: How Corynebacterium Diphtheriae Causes Illnessmentioning

confidence: 99%

“…Diphtheria toxin is translocated by receptormediated endocytosis when subunit B binds to its specific cell-surface receptor, which is a receptor for heparin-binding epidermal growth factor (EGF)-like growth factor [81,84,90]. The presence or absence of this receptor is dependent on the cell type [54,57,81]. Once the diphtheria toxin has entered the endosome, the disulfide bond connecting subunits A and B is reduced and subunit A is released into the cytosol [60].…”

Section: How Corynebacterium Diphtheriae Causes Illnessmentioning

confidence: 99%

The Evolution and Value of Diphtheria Vaccine

Bae¹

2011

KSBB Journal

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1) This review article provides an overview of the evolution of diphtheria vaccine, its value and its future. Diphtheria is an infectious illness caused by diphtheria toxin produced by pathogenic strains of Corynebacterium diphtheriae. It is characterized by a sore throat with membrane formation due to local tissue necrosis, which can lead to fatal airway obstruction; neural and cardiac damage are other common complications. Diphtheria vaccine was first brought to market in the 1920s, following the discovery that diphtheria toxin can be detoxified using formalin. However, conventional formalin-inactivated toxoid vaccines have some fundamental limitations. Innovative technologies and approaches with the potential to overcome these limitations are discussed in this paper. These include genetic inactivation of diphtheria toxoid, innovative vaccine delivery systems, new adjuvants (both TLRindependent and TLR-dependent adjuvants), and heat-and freeze-stable agents, as well as novel platforms for producing improved conventional vaccine, DNA vaccine, transcutaneous (microneedle-mediated) vaccine, oral vaccine and edible vaccine expressed in transgenic plants. These innovations target improvements in vaccine quality (efficacy, safety, stability and consistency), ease of use and/or thermal stability. Their successful development and use should help to increase global diphtheria vaccine coverage.

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Structure-Function Analysis of the Diphtheria Toxin Receptor Toxin Binding Site by Site-directed Mutagenesis

Cited by 47 publications

References 41 publications

Suppression of the Biological Activities of the Epidermal Growth Factor (EGF)-like Domain by the Heparin-binding Domain of Heparin-binding EGF-like Growth Factor

Suppression of the Biological Activities of the Epidermal Growth Factor (EGF)-like Domain by the Heparin-binding Domain of Heparin-binding EGF-like Growth Factor

Role of EGF Receptor Activation in Angiotensin II–Induced Renal Epithelial Cell Hypertrophy

The Evolution and Value of Diphtheria Vaccine

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