title: ERp57 is essential for efficient folding of glycoproteins sharing common structural domains.
abstract: ERp57 is a member of the protein disulphide isomerase family of oxidoreductases, which are involved in native disulphide bond formation in the endoplasmic reticulum of mammalian cells. This enzyme has been shown to be associated with both calnexin and calreticulin and, therefore, has been proposed to be a glycoprotein-specific oxidoreductase. Here, we identify endogenous substrates for ERp57 by trapping mixed disulphide intermediates between enzyme and substrate. Our results demonstrate that the substrates for this enzyme are mostly heavily glycosylated, disulphide bonded proteins. In addition, we show that the substrate proteins share common structural domains, indicating that substrate specificity may involve specific structural features as well as the presence of an oligosaccharide side chain. We also show that the folding of two of the endogenous substrates for ERp57 is impaired in ERp57 knockout cells and that prevention of an interaction with calnexin or calreticulin perturbs the folding of some, but not all, substrates with multiple disulphide bonds. These results suggest a specific role for ERp57 in the isomerisation of non-native disulphide bonds in specific glycoprotein substrates.
authors.affiliation: Faculty of Life Sciences, University of Manchester, Manchester, UK.
authors.forename: Catherine E
authors.initials: CE
authors.lastname: Jessop
authors.forename: Seema
authors.initials: S
authors.lastname: Chakravarthi
authors.forename: Natalio
authors.initials: N
authors.lastname: Garbi
authors.forename: Günter J
authors.initials: GJ
authors.lastname: Hämmerling
authors.forename: Simon
authors.initials: S
authors.lastname: Lovell
authors.forename: Neil J
authors.initials: NJ
authors.lastname: Bulleid
mesh_terms: D000818:Animals; D037281:Calnexin / Q000737:chemistry; D037282:Calreticulin / Q000737:chemistry; D051152:Clusterin / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D010100:Oxygen / Q000737:chemistry; D011485:Protein Binding; D019704:Protein Disulfide-Isomerases / Q000737:chemistry / Q000502:physiology*; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011817:Rabbits; D013379:Substrate Specificity
chemical_list: D037282:Calreticulin; D051152:Clusterin; D004220:Disulfides; D006023:Glycoproteins; D037281:Calnexin; D019704:Protein Disulfide-Isomerases; C085868:PDIA3 protein, human; D010100:Oxygen