Conformational changes in plant toxins

CHelnokova OV

Conformational changes in plant toxins during intracellular transport

03.00.04 - biochemistry

Diss. ... Kand.biol.nauk

2002


Table of Contents.

Abbreviations

Introduction

1.1. Ricin and viscumin - representatives of vegetable RIB type II

1.1.1. The structure of the catalytic subunit of ricin and viscumin

1.1.2. The structure of the binding subunits of ricin and viscumin

1.1.3. The interaction between subunits of ricin and viscumin

1.2. Intracellular transport protein toxins

1.2.1. Binding to the membrane: receptors for ricin and viscumin

1.2.2. Internalization of toxin into the cell

1.2.3. Vesicular transport proteins in the cell

1.2.4. Passage of toxins through the endosomal compartments cells

1.2.5. Transport toxin to the Golgi

1.2.6. Retrograde transport of toxins in the endoplasmic reticulum

1.2.7. Translocation of toxins

Closing

Chapter 2. Materials and methods

2.1. Cages

2.2. Animals

2.3. Isolation of ricin and its subunits from the plant Ricinus communis

2.4. Isolation of toxin and subunits of plants Viscum album

2.5. Preparation of hybridomas producing antibodies against the A subunit of ricin

2.6. Preparation of hybridomas producing antibodies against the B subunit viscumin

2.7. Antibody Production and Purification

2.8. Determination of antibody isotype

2.9. Biotinylation of proteins

2.10. Study of the interaction of the antibody with the antigen by enzyme-linked immunosorbent assay

2.11. Competitive Enzyme Immunoassay

2.12. Sandwich ELISA Based on the MoAbs

2.13. Evaluation of galactose-binding activity of toxins

2.14. Dissociation of the subunits of ricin and viscumin under the influence of DTT

2.15. Influence of mAbs against MLB to interact viscumin and the B chain with asialofetuin

2.16. Cell ELISA

2.17. Study of recycling viscumin cells TA71

2.18. Effect of exposure to guanidine hydrochloride epitope recognized by antibody TA71

2.19. The peptide ELISA

2.20. Evaluation of cytotoxic activity of toxins

2.21. Acellular system of translation

2.22. The electrophoresis system of Laemmli

2.23. Immunoblotting

Chapter 3. Results and Discussion

3.1. The study of the mechanism of action of ricin and viscumin stage intracellular dissociation

3.2. The inhibitory effect on viscumin hybridomas that synthesize antibodies against its binding subunit

3.3. A conformational change in the subunit viscumin intracellular translocation

Findings

References