The essential function of yeast protein disulfide isomerase does not reside in its isomerase activity.
Protein disulfide isomerase (PDI) is believed to function in vivo by catalyzing the isomerization of disulfide bonds in proteins and thereby facilitating their folding. In S. cerevisiae PDI is encoded by an essential gene. Deletion of nearly one-third of the C-terminal residues of PDI altered PDI's cellular localization but not cell viability. Further deletions resulted in lethality, but these truncated proteins still exhibited PDI activity in vitro. Cells carrying a variant PDI in which both-CGHC-active sites were disrupted were viable. However, these cells exhibited a delay in the disulfide bond formation and transport of carboxypeptidase Y. In vitro enzyme assays revealed that disruption of both sites abolished catalytic activity. These results indicate that PDI catalyzes disulfide bond formation both in vivo and in vitro and that the integrity of the active sites is required for catalysis. However, this catalytic activity is not essential for yeast viability.
Department of Biochemistry and Cell Biology, State University of New York at Stony Brook 11794-5215.
M L
ML
LaMantia
W J
WJ
Lennarz
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004220:Disulfides / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005800:Genes, Fungal; D005982:Glutathione Transferase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007535:Isomerases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D016133:Polymerase Chain Reaction; D019704:Protein Disulfide-Isomerases; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D017384:Sequence Deletion; D013379:Substrate Specificity
D004220:Disulfides; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D007535:Isomerases; D019704:Protein Disulfide-Isomerases