Instability of toxin A subunit of AB5 toxins in the bacterial periplasm caused by deficiency of their cognate B subunits

• Published in: Biochimica et biophysica acta 2011-10, Vol.1808 (10), p.2359-2365
• Main Authors: Kim, Sang-Hyun, Ryu, Su Hyang, Lee, Sang-Ho, Lee, Yong-Hoon, Lee, Sang-Rae, Huh, Jae-Won, Kim, Sun-Uk, Kim, Ekyune, Kim, Sunghyun, Jon, Sangyong, Bishop, Russell E., Chang, Kyu-Tae
• Format: Article
• Language: English
• Subjects: Escherichia coli; Heat-labile toxin; Outer membrane; Periplasm; Shiga toxin
• ISSN: 0005-2736; 0006-3002; 1879-2642; 1878-2434
• DOI: 10.1016/j.bbamem.2011.06.016

Shiga toxin (STx) belongs to the AB5 toxin family and is transiently localized in the periplasm before secretion into the extracellular milieu. While producing outer membrane vesicles (OMVs) containing only A subunit of the toxin (STxA), we created specific STx1B- and STx2B-deficient mutants of E. coli O157:H7. Surprisingly, STxA subunit was absent in the OMVs and periplasm of the STxB-deficient mutants. In parallel, the A subunit of heat-labile toxin (LT) of enterotoxigenic E. coli (ETEC) was absent in the periplasm of the LT-B-deficient mutant, suggesting that instability of toxin A subunit in the absence of the B subunit is a common phenomenon in the AB5 bacterial toxins. Moreover, STx2A was barely detectable in the periplasm of E. coli JM109 when stx2A was overexpressed alone, while it was stably present when stxB was co-expressed. Compared with STx2 holotoxin, purified STx2A was degraded rapidly by periplasmic proteases when assessed for in vitro proteolytic susceptibility, suggesting that the B subunit contributes to stability of the toxin A subunit in the periplasm. We propose a novel role for toxin B subunits of AB5 toxins in protection of the A subunit from proteolysis during holotoxin assembly in the periplasm. ► We expressed the A subunit of Shiga toxin by itself in the bacterial periplasm. ► We found that its stability depended on co-expression of its cognate B subunit. ► We conclude that the B subunit protects the A subunit from periplasmic proteases.