title: Experimental and theoretical analyses of the domain architecture of mammalian protein disulphide-isomerase.
abstract: The high resolution structure of full-length protein disulphide-isomerase (PDI) has not been determined, but the polypeptide is generally assumed to comprise a series of consecutive domains. Models of its domain organisation have been proposed on the basis of various sequence-based criteria and, more recently, from structural studies on recombinant fragments corresponding to putative domains. We here describe direct studies of the domain architecture of full-length mammalian PDI based on limited proteolysis of the native enzyme. The results are consistent with an emerging model based on the existence of 4 consecutive domains each with the thioredoxin fold. The model was further tested by expressing recombinant fragments corresponding to alternative domain models and to truncated domains; the observed properties of these purified fragments supported the 4-domain model. A multiple alignment of many PDI-like sequences was generated to test whether domain boundaries could be predicted from any features of the alignment, such as sequence variability or hydrophilicity; neither of these parameters reliably predicted the domain boundaries determined by experiment.
authors.affiliation: Research School of Biosciences, University of Kent, Canterbury, UK.
authors.forename: R B
authors.initials: RB
authors.lastname: Freedman
authors.forename: P J
authors.initials: PJ
authors.lastname: Gane
authors.forename: H C
authors.initials: HC
authors.lastname: Hawkins
authors.forename: R
authors.initials: R
authors.lastname: Hlodan
authors.forename: S H
authors.initials: SH
authors.lastname: McLaughlin
authors.forename: J W
authors.initials: JW
authors.lastname: Parry
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003001:Cloning, Molecular; D006868:Hydrolysis; D008099:Liver / Q000201:enzymology; D008956:Models, Chemical; D019704:Protein Disulfide-Isomerases / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary
chemical_list: D019704:Protein Disulfide-Isomerases