Sep 11, 2002

Clostridium perfringens iota toxin. Mapping of the Ia domain involved in docking with Ib and cellular internalization

The Journal of Biological Chemistry
J C MarvaudMichel R Popoff

Abstract

Clostridium perfringens iota toxin consists of two unlinked proteins. The binding component (Ib) is required to internalize into cells an enzymatic component (Ia) that ADP-ribosylates G-actin. To characterize the Ia domain that interacts with Ib, fusion proteins were constructed between the C. botulinum C3 enzyme, which ADP-ribosylates Rho, and various truncated versions of Ia. These chimeric molecules retained the wild type ADP-ribosyltransferase activity specific for Rho and were recognized by antibodies against C3 enzyme and Ia. Internalization of each chimera into Vero cells was assessed by measuring the disorganization of the actin cytoskeleton and intracellular ADP-ribosylation of Rho. Fusion proteins containing C3 linked to the C terminus of Ia were transported most efficiently into cells like wild type Ia in an Ib-dependent manner that was blocked by bafilomycin A1. The minimal Ia fragment that promoted translocation of Ia-C3 chimeras into cells consisted of 128 central residues (129-257). These findings revealed that iota toxin is a suitable system for mediating the entry of heterologous proteins such as C3 into cells.

  • References29
  • Citations25

Citations

Mentioned in this Paper

Flow Cytometry
Immunoblotting, Reverse
Botulinum Toxins
Alkalescens-Dispar Group
Chimeric Proteins, Recombinant
Isoactin
Toxin
Bacterial Toxins
SDS-PAGE
Protoplasm

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