Flexibility in the receptor-binding domain of the enzymatic colicin E9 is required for toxicity against Escherichia coli cells

Journal of Bacteriology
Christopher N PenfoldR James

Abstract

The events that occur after the binding of the enzymatic E colicins to Escherichia coli BtuB receptors that lead to translocation of the cytotoxic domain into the periplasmic space and, ultimately, cell killing are poorly understood. It has been suggested that unfolding of the coiled-coil BtuB receptor binding domain of the E colicins may be an essential step that leads to the loss of immunity protein from the colicin and immunity protein complex and then triggers the events of translocation. We introduced pairs of cysteine mutations into the receptor binding domain of colicin E9 (ColE9) that resulted in the formation of a disulfide bond located near the middle or the top of the R domain. After dithiothreitol reduction, the ColE9 protein with the mutations L359C and F412C (ColE9 L359C-F412C) and the ColE9 protein with the mutations Y324C and L447C (ColE9 Y324C-L447C) were slightly less active than equivalent concentrations of ColE9. On oxidation with diamide, no significant biological activity was seen with the ColE9 L359C-F412C and the ColE9 Y324C-L447C mutant proteins; however diamide had no effect on the activity of ColE9. The presence of a disulfide bond was confirmed in both of the oxidized, mutant proteins by matrix-assis...Continue Reading

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Citations

Nov 17, 2011·Quarterly Reviews of Biophysics·Grigorios PapadakosColin Kleanthous
May 18, 2012·The Journal of Biological Chemistry·Silke I PatzerKornelius Zeth
May 22, 2009·The Journal of Biological Chemistry·Mireille VankemmelbekeRichard James
Feb 6, 2007·Journal of Bacteriology·Muriel MasiRajeev Misra
Sep 17, 2005·Journal of Bacteriology·Sarah L HandsChristopher N Penfold
Mar 10, 2007·Microbiology and Molecular Biology Reviews : MMBR·Eric CascalesDanièle Cavard
Sep 17, 2005·Proceedings of the National Academy of Sciences of the United States of America·Nicholas G HousdenColin Kleanthous
Aug 22, 2009·The EMBO Journal·Daniel A BonsorColin Kleanthous
May 16, 2013·Molecular Microbiology·Christopher L JohnsonJeremy H Lakey
Sep 17, 2013·MicrobiologyOpen·Mireille VankemmelbekeChristopher N Penfold
Nov 28, 2012·Biochemical Society Transactions·Nicholas G Housden, Colin Kleanthous
Apr 22, 2014·Biochimica Et Biophysica Acta·Young Chan KimChristopher N Penfold
Oct 28, 2005·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Aug 28, 2007·The Journal of Biological Chemistry·Daniel WalkerColin Kleanthous
Jul 13, 2016·The Biochemical Journal·Alexander KleinColin Kleanthous
Apr 20, 2019·Frontiers in Microbiology·Iva Atanaskovic, Colin Kleanthous

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