Diphtheria Toxin, CRM197 Mutant

Cat.No: FNK-01-515

Size: 200 μg

Storage: -80℃. Avoid freeze-thaw cycles.

Concentration: 1.0 mg/ml

Buffer: 20 mM Tris-HCl (pH 7.2), 150 mM NaCl, 10% Glycerol

Purity: More than 95% purity (see below; SDS-PAGE without 2-mercaptoethanol) 

Speical Note: For resaech use only, not for human use.
 

Product Description

CRM197 was highly purified from growth media of the Corynebacterium diphtheriae mutant as mostly unnicked form.

Biochemical Activity

Nontoxic mutant of Diphtheria toxin.

While CRM197 shows no enzymatic activity, it is immunologically indistinguishable from wild-type Diphtheria toxin. 

Application 

1.CRM197 retains activity to bind the receptor, HB-EGF (Heparin-Binding EGF-like Growth Factor) and inhibits the growth-stimulating activity of  HB-EGF (Ref.1)

2. Putative drug for treatment of malignant tumors such as ovarian tumor, which secretes higher levels of HB-EGF (Ref 2).

3. Western blotting 

4. SDS-PAGE

5. ELISA 

Background

CRM197 (Cross-Reacting Material 197) is a nontoxic mutant of Diphtheria toxin. CRM197, like wild-type Diphtheria toxin, is composed of a single polypeptide chain of 535 amino acids (58 kD) and nicked by cellular protease like furin to give fragments A (N-terminal, 21 kDa) and B (C-terminal, 37 kDa) which are linked by disulfide bridges. Binding to the cell surface of fragment B allows fragment A to penetrate the host cell. Fragment A of wild-type toxin catalyzes the ADPribosylation of eucaryotic elongation factor-2 (eEF2) by using NAD as a substrate, thus inactivating eEF2 and inhibiting protein synthesis. However, CRM197 has an alteration of 52nd Gly to Glu and has neither ADP ribosylation activity nor toxicity to cells. While CRM197 shows no enzymatic activity, it is immunologically indistinguishable from wild-type Diphteria toxin. CRM197 competitively inhibits binding of HB-EGF to HB-EGF receptor which is also Diphtheria toxin receptor.

Data Link

UniProtKB Q5PY51

Please Note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. NOT FOR MILITRAY USE.

Date Image

01-515 Diphtheria Toxin Mutant CRM197, functional 



Fig.1 SDS-PAGE of Diphtheria Toxin Mutant CRM197 

Referneces

This product was used in follwoing Ref. 

1. Mitamura T. et al. “Structure-function analysis of thediphtheria toxin receptor toxin binding site by site-directed mutagenesis.” J. Biol. Chem.272: 27084-27090 (1997) PMID: 9341148

2. Miyamoto S. et al. “Heparin-binding EGF-like growth factor is a promising target for ovarian cancer therapy.” Cancer Res. 64: 5720-5727 (2004) PMID: 15313912