Antibodies Against Recombinant Shiga Toxin Subunit B Neutralize Shiga Toxin Toxicity in HeLa Cells
Authors: Gupta, Pallavi; Kumar Singh, Manglesh; Singh, Padma; Tiwari, Mugdha; Kumar Dhaked, Ram
Source: Protein and Peptide Letters, Volume 17, Number 6, June 2010 , pp. 774-781(8)
Publisher: Bentham Science Publishers
Abstract:Shigella dysenteriae type 1 and Escherichia coli O157:H7 produce Shiga toxin (Stx) and Shiga toxin (Stx1), respectively and these two toxins are almost identical. E. coli O157:H7 is the major cause of diarrhea-associated hemolytic uremic syndrome. Stx and Stx1 are AB5 type of toxin with a molecular weight of 70 kDa, comprising an enzymaticalyactive A subunit (32 kDa) and five receptor-binding B subunits (7.7 kDa). In this study DNA fragment (289 bp, Gene Bank Accn No. EF685161) coding for B chain of Stx was amplified from S. dysenteriae type1 and cloned. Shiga toxin-binding subunit was expressed and purified in native conditions by affinity and gel permeation chromatography with the yield of 5.1 mg/L in shake flask culture. For the purpose of immunization, the polypeptide was polymerized with glutaraldehyde. Hyper immune serum produced in mice reacted with the purified polypeptide and intact Shiga toxin. The anti- StxB antiserum effectively neutralized the cytotoxicity of Shiga toxin towards HeLa cells.
Document Type: Research Article
Publication date: June 2010
- Protein & Peptide Letters publishes short papers in all important aspects of protein and peptide research, including structural studies, recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, drug design etc. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallisation, and preliminary structure determinations of biologically important proteins are acceptable. Purely theoretical papers are also acceptable provided they provide new insight into the principles of protein/peptide structure and function.