title: Expression, purification, refolding and crystallization of the carbohydrate-recognition domain of p58/ERGIC-53, an animal C-type lectin involved in export of glycoproteins from the endoplasmic reticulum.
abstract: p58/ERGIC-53 is a mammalian calcium-dependent lectin that serves as a glycoprotein-sorting receptor between the endoplasmic reticulum (ER) and the Golgi complex. It is a type I transmembrane protein with two lumenal domains, one of which is a carbohydrate-recognition domain (CRD) and homologous to leguminous lectins. The CRD of p58, the rat homologue of human ERGIC-53, was overexpressed in insect cells and Escherichia coli, purified and crystallized using Li(2)SO(4) as a precipitant. The crystals belong to space group I222, with unit-cell parameters a = 49.6, b = 86.1, c = 128.1 A, and contain one molecule per asymmetric unit, corresponding to a packing density of 2.4 A(3)Da(-1). Knowledge of the structure of p58/ERGIC-53 will provide a starting model for understanding receptor-mediated glycoprotein sorting between the ER and the Golgi.
authors.affiliation: Molecular Structural Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-17177 Stockholm, Sweden.
authors.forename: Lucas M
authors.initials: LM
authors.lastname: Velloso
authors.forename: Kerstin
authors.initials: K
authors.lastname: Svensson
authors.forename: Ulla
authors.initials: U
authors.lastname: Lahtinen
authors.forename: Gunter
authors.initials: G
authors.lastname: Schneider
authors.forename: Ralf F
authors.initials: RF
authors.lastname: Pettersson
authors.forename: Ylva
authors.initials: Y
authors.lastname: Lindqvist
mesh_terms: D000818:Animals; D002241:Carbohydrates / Q000737:chemistry; D003460:Crystallization; D018360:Crystallography, X-Ray; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry*; D011487:Protein Conformation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D051381:Rats
chemical_list: D002241:Carbohydrates; D006023:Glycoproteins; C084004:LMAN1 protein, human; D037102:Lectins; C501458:Lman1 protein, rat; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins