title: The pertussis toxin S1 subunit is a thermally unstable protein susceptible to degradation by the 20S proteasome.
abstract: Pertussis toxin (PT) is an AB-type protein toxin that consists of a catalytic A subunit (PT S1) and an oligomeric, cell-binding B subunit. It belongs to a subset of AB toxins that move from the cell surface to the endoplasmic reticulum (ER) before the A chain passes into the cytosol. Toxin translocation is thought to involve A chain unfolding in the ER and the quality control mechanism of ER-associated degradation (ERAD). The absence of lysine residues in PT S1 may allow the translocated toxin to avoid ubiquitin-dependent degradation by the 26S proteasome, which is the usual fate of exported ERAD substrates. As the conformation of PT S1 appears to play an important role in toxin translocation, we used biophysical and biochemical methods to examine the structural properties of PT S1. Our in vitro studies found that the isolated PT S1 subunit is a thermally unstable protein that can be degraded in a ubiquitin-independent fashion by the core 20S proteasome. The thermal denaturation of PT S1 was inhibited by its interaction with NAD, a donor molecule used by PT S1 for the ADP ribosylation of target G proteins. These observations support a model of intoxication in which toxin translocation, degradation, and activity are all influenced by the heat-labile nature of the isolated toxin A chain.
authors.affiliation: Department of Molecular Biology and Microbiology, University of Central Florida, 12722 Research Parkway, Orlando, Florida 32826, USA.
authors.forename: Abhay H
authors.initials: AH
authors.lastname: Pande
authors.forename: David
authors.initials: D
authors.lastname: Moe
authors.forename: Maneesha
authors.initials: M
authors.lastname: Jamnadas
authors.forename: Suren A
authors.initials: SA
authors.lastname: Tatulian
authors.forename: Ken
authors.initials: K
authors.lastname: Teter
mesh_terms: D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D006868:Hydrolysis; D037342:Pertussis Toxin / Q000737:chemistry / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D011489:Protein Denaturation; D013056:Spectrophotometry, Ultraviolet; D013501:Surface-Active Agents / Q000737:chemistry; D013816:Thermodynamics
chemical_list: D013501:Surface-Active Agents; C083388:pertussis toxin, S1 subunit; D037342:Pertussis Toxin; D046988:Proteasome Endopeptidase Complex