Sulfhydryl oxidation, not disulfide isomerization, is the principal function of protein disulfide isomerase in yeast Saccharomyces cerevisiae.
Protein disulfide isomerase (PDI) is an essential protein folding assistant of the eukaryotic endoplasmic reticulum that catalyzes both the formation of disulfides during protein folding (oxidase activity) and the isomerization of disulfides that may form incorrectly (isomerase activity). Catalysis of thiol-disulfide exchange by PDI is required for cell viability in Saccharomyces cerevisiae, but there has been some uncertainty as to whether the essential role of PDI in the cell is oxidase or isomerase. We have studied the ability of PDI constructs with high oxidase activity and very low isomerase activity to complement the chromosomal deletion of PDI1 in S. cerevisiae. A single catalytic domain of yeast PDI (PDIa') has 50% of the oxidase activity but only 5% of the isomerase activity of wild-type PDI in vitro. Titrating the expression of PDI using the inducible/repressible GAL1-10 promoter shows that the amount of wild-type PDI protein needed to sustain a normal growth rate is 60% or more of the amount normally expressed from the PDI1 chromosomal location. A single catalytic domain (PDIa') is needed in molar amounts that are approximately twice as high as those required for wild-type PDI, which contains two catalytic domains. This comparison suggests that high (>60%) PDI oxidase activity is critical to yeast growth and viability, whereas less than 6% of its isomerase activity is needed.
Solovyov
Anton
A
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA.
Xiao
Ruoyu
R
Gilbert
Hiram F
HF
D000818:Animals; D002384:Catalysis; D020134:Catalytic Domain; D043402:Cathepsin A / Q000378:metabolism; D002455:Cell Division; D002470:Cell Survival; D002875:Chromosomes / Q000378:metabolism; D004220:Disulfides; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D017353:Gene Deletion; D005816:Genetic Complementation Test; D005947:Glucose / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D019704:Protein Disulfide-Isomerases / Q000737:chemistry* / Q000502:physiology; D017434:Protein Structure, Tertiary; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013439:Sulfhydryl Reagents / Q000737:chemistry*; D013997:Time Factors
D004220:Disulfides; D013439:Sulfhydryl Reagents; D043402:Cathepsin A; D019704:Protein Disulfide-Isomerases; D005947:Glucose; D010100:Oxygen