PMID: 8609372Feb 1, 1996Paper

Cytotoxicity of KDEL-terminated ricin toxins correlates with distribution of the KDEL receptor in the Golgi

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
E TaggeA Frankel

Abstract

DNAs encoding ricin toxin A chain (RTA), with or without a C-terminal endoplasmic reticulum retention signal KDEL, were subcloned into pGEX2T bacterial expression plasmid. After transformation of JM105 E. coli cells and induction with isopropylthio-beta-galactoside (IPTG), fusion proteins were bound to an immobilized glutathione matrix and recombinant ricin A chains released with thrombin. Both recombinant wild-type RTA and RTA with KDEL had immunological reactivity and catalytic activity indistinguishable from plant RTA. The bacterial RTA products reassociated with plant ricin B chain (RTB) similarly to plant RTA. Cell cytotoxicities were measured on seven cell lines for each A-chain and heterodimer. Although KDEL sequences enhanced cytotoxicity in most cases, significant variability was observed. In each case, addition of KDEL enhanced A-chain cytotoxicity more than holotoxin cytotoxicity. Three cell lines showed reduced KDEL enhancement of both RTA and ricin cytotoxicity. The concentration of KDEL receptor was examined on each cell line by immunofluorescence microscopy with an antireceptor monoclonal antibody. Differences in sensitivity to KDEL-containing toxins correlated with altered distribution of KDEL receptor between e...Continue Reading

Citations

Oct 12, 2001·Toxicon : Official Journal of the International Society on Toxinology·S Olsnes, J V Kozlov
Dec 17, 2002·Journal of Cell Science·Marcia C Haigis, Ronald T Raines
Apr 15, 1997·Proceedings of the National Academy of Sciences of the United States of America·A RapakS Olsnes
Feb 5, 2003·Biochemical and Biophysical Research Communications·Gunaratna Kuttuva RajaraoLiam Good
Aug 11, 2004·Toxicon : Official Journal of the International Society on Toxinology·Sjur Olsnes
Jan 26, 2011·Journal of Molecular Biology·Morten H H NørholmGunnar von Heijne
Jun 23, 2009·BMC Cell Biology·Joseph T ByersBalwant S Tuana
Sep 10, 1998·Biochemical and Biophysical Research Communications·M SvinthJ D Robertus
May 16, 1997·Carbohydrate Research·A E FrankelM C Willingham

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