title: Novel protein-disulfide isomerases from the early-diverging protist Giardia lamblia.
abstract: Protein-disulfide isomerase is essential for formation and reshuffling of disulfide bonds during nascent protein folding in the endoplasmic reticulum. The two thioredoxin-like active sites catalyze a variety of thiol-disulfide exchange reactions. We have characterized three novel protein-disulfide isomerases from the primitive eukaryote Giardia lamblia. Unlike other protein-disulfide isomerases, the giardial enzymes have only one active site. The active-site sequence motif in the giardial proteins (CGHC) is characteristic of eukaryotic protein-disulfide isomerases, and not other members of the thioredoxin superfamily that have one active site, such as thioredoxin and Dsb proteins from Gram-negative bacteria. The three giardial proteins have very different amino acid sequences and molecular masses (26, 50, and 13 kDa). All three enzymes were capable of rearranging disulfide bonds, and giardial protein-disulfide isomerase-2 also displayed oxidant and reductant activities. Surprisingly, the three giardial proteins also had Ca(2+)-dependent transglutaminase activity. This is the first report of protein-disulfide isomerases with a single active site that have diverse roles in protein cross-linking. This study may provide clues to the evolution of key functions of the endoplasmic reticulum in eukaryotic cells, protein disulfide formation, and isomerization.
authors.affiliation: Department of Pathology, Division of Infectious Diseases, University of California, San Diego, California 92103-8416, USA.
authors.forename: L A
authors.initials: LA
authors.lastname: Knodler
authors.forename: R
authors.initials: R
authors.lastname: Noiva
authors.forename: K
authors.initials: K
authors.lastname: Mehta
authors.forename: J M
authors.initials: JM
authors.lastname: McCaffery
authors.forename: S B
authors.initials: SB
authors.lastname: Aley
authors.forename: S G
authors.initials: SG
authors.lastname: Svärd
authors.forename: T G
authors.initials: TG
authors.lastname: Nystul
authors.forename: D S
authors.initials: DS
authors.lastname: Reiner
authors.forename: J D
authors.initials: JD
authors.lastname: Silberman
authors.forename: F D
authors.initials: FD
authors.lastname: Gillin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan; D004721:Endoplasmic Reticulum / Q000201:enzymology / Q000648:ultrastructure; D016829:Giardia lamblia / Q000201:enzymology*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D019704:Protein Disulfide-Isomerases / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D011503:Transglutaminases / Q000378:metabolism
chemical_list: D016054:DNA, Protozoan; D011503:Transglutaminases; D019704:Protein Disulfide-Isomerases