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.
Keywords: S. dysenteriae type1, Shiga toxin, Shiga like toxin, cytotoxicity, hemolytic uremic syndrome (HUS)
Protein & Peptide Letters
Title: Antibodies Against Recombinant Shiga Toxin Subunit B Neutralize Shiga Toxin Toxicity in HeLa Cells
Volume: 17 Issue: 6
Author(s): Pallavi Gupta, Manglesh Kumar Singh, Padma Singh, Mugdha Tiwari and Ram Kumar Dhaked
Affiliation:
Keywords: S. dysenteriae type1, Shiga toxin, Shiga like toxin, cytotoxicity, hemolytic uremic syndrome (HUS)
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.
Export Options
About this article
Cite this article as:
Gupta Pallavi, Kumar Singh Manglesh, Singh Padma, Tiwari Mugdha and Kumar Dhaked Ram, Antibodies Against Recombinant Shiga Toxin Subunit B Neutralize Shiga Toxin Toxicity in HeLa Cells, Protein & Peptide Letters 2010; 17 (6) . https://dx.doi.org/10.2174/092986610791190291
DOI https://dx.doi.org/10.2174/092986610791190291 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Oncologists Current Opinion on the Treatment of Colon Carcinoma
Anti-Cancer Agents in Medicinal Chemistry Neonatal Fever in the Term Infant: Evaluation and Management Strategies
Current Pediatric Reviews Inflammatory Responses in Brain Ischemia
Current Medicinal Chemistry Specific Recognition of DNA by Small Molecules
Current Medicinal Chemistry Synthesis and Preliminary COX-2 Expression Inhibitory Activities of Andrographolide Derivatives
Letters in Drug Design & Discovery Hyperglycemia Induced Changes in Liver: In vivo and In vitro Studies
Current Diabetes Reviews Importance of Wnt Signaling in the Tumor Stroma Microenvironment
Current Cancer Drug Targets Hyperhomocysteinaemia: A Critical Review of Old and New Aspects
Current Drug Metabolism Development of Radioligands for In Vivo Imaging of Type 1 Cannabinoid Receptors (CB1) in Human Brain
Current Pharmaceutical Design Immunization with Peptide-Protein Conjugates: Impact on Benchmarking B-Cell Epitope Prediction for Vaccine Design
Protein & Peptide Letters Bacterial Toxins: Potential Weapons Against HIV Infection
Current Pharmaceutical Design Testicular Germ Cell Tumors: A Paradigm for the Successful Treatment of Solid Tumor Stem Cells
Current Cancer Therapy Reviews Pathobiology and Prevention of Cancer Chemotherapy-Induced Bone Growth Arrest, Bone Loss, and Osteonecrosis
Current Molecular Medicine Expression, Purification and Immunological Characterization of Recombinant Shiga Toxin A Subunit
Protein & Peptide Letters Role of Wnt Signaling in Tissue Fibrosis, Lessons from Skeletal Muscle and Kidney
Current Molecular Medicine Pharmacokinetic Mechanisms for Reduced Toxicity of Irinotecan by Coadministered Thalidomide
Current Drug Metabolism Rallying the Troops: Innate Mucosal Responses to Enteric Pathogens
Current Immunology Reviews (Discontinued) New Trends in Medicinal Chemistry Approaches to Antiobesity Therapy
Current Topics in Medicinal Chemistry Myocardial Perfusion and Coronary Vasomotor Function: Emerging Role of PET Imaging
Vascular Disease Prevention (Discontinued) Emerging Treatments in Acute Lymphoblastic Leukemia
Current Cancer Drug Targets