Abbreviated Journal Title
J. Biol. Chem.
RETICULUM-ASSOCIATED DEGRADATION; INTESTINAL MICROVILLUS MEMBRANE; CHO-CELLS RESISTANT; ENDOPLASMIC-RETICULUM; A(1) DOMAIN; RETRO-TRANSLOCATION; SIGNAL-TRANSDUCTION; THERMAL-STABILITY; SHIGA; TOXIN; C-TERMINUS; Biochemistry & Molecular Biology
Cholera toxin (CT) travels from the cell surface to the endoplasmic reticulum (ER) as an AB holotoxin. ER-specific conditions then promote the dissociation of the catalytic CTA1 subunit from the rest of the toxin. CTA1 is held in a stable conformation by its assembly in the CT holotoxin, but the dissociated CTA1 subunit is an unstable protein that spontaneously assumes a disordered state at physiological temperature. This unfolding event triggers the ER-to-cytosol translocation of CTA1 through the quality control mechanism of ER-associated degradation. The translocated pool of CTA1 must regain a folded, active structure to modify its G protein target which is located in lipid rafts at the cytoplasmic face of the plasma membrane. Here, we report that lipid rafts place disordered CTA1 in a functional conformation. The hydrophobic C-terminal domain of CTA1 is essential for binding to the plasma membrane and lipid rafts. These interactions inhibit the temperature-induced unfolding of CTA1. Moreover, lipid rafts could promote a gain of structure in the disordered, 37 degrees C conformation of CTA1. This gain of structure corresponded to a gain of function: whereas CTA1 by itself exhibited minimal in vitro activity at 37 degrees C, exposure to lipid rafts resulted in substantial toxin activity at 37 degrees C. In vivo, the disruption of lipid rafts with filipin substantially reduced the activity of cytosolic CTA1. Lipid rafts thus exhibit a chaperone-like function that returns disordered CTA1 to an active state and is required for the optimal in vivo activity of CTA1.
Journal of Biological Chemistry
Ray, Supriyo; Taylor, Michael; Banerjee, Tuhina; Tatulian, Suren A.; and Teter, Ken, "Lipid Rafts Alter the Stability and Activity of the Cholera Toxin A1 Subunit" (2012). Faculty Bibliography 2010s. 3176.