Catalysis of the oxidative folding of ribonuclease A by protein disulfide isomerase: pre-steady-state kinetics and the utilization of the oxidizing equivalents of the isomerase

Tunicamycin-inducible gene A polypeptide (TIGA) is a member of the protein disulfide isomerase (PDI) family and is suggested to facilitate the folding of nascent polypeptides. The functional properties of TIGA were investigated here. TIGA acted as an isomerase, catalyzing the refolding of denatured and reduced ribonuclease A. TIGA also exhibited chaperone activity in the refolding of denatured prochymosin but not in the refolding of glyceraldehyde 3-phosphate dehydrogenase (GAPDH), indicating that it had substrate specificity with respect to chaperone activity. Detailed study with a series of thioredoxin-motif (trx-motif) mutants revealed that the 2 trx-motifs of TIGA were not equal in activity. The N-terminal trx-motif was more active than the C-terminal trx-motif, and the first cysteine in each trx-motif was necessary for isomerase activity.Key words: tunicamycin-inducible gene A polypeptide (TIGA), protein disulfide isomerase, chaperone, protein refolding.

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Faculty Opinions recommendation of Dynamic assembly of protein disulfide isomerase in catalysis of oxidative folding.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.735573116.793560095 ◽

2019 ◽

Author(s):

Robert Flaumenhaft

Keyword(s):

Protein Disulfide Isomerase ◽

Oxidative Folding ◽

Disulfide Isomerase ◽

Dynamic Assembly ◽

Protein Disulfide

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Mutations in domaina′ of protein disulfide isomerase affect the folding pathway of bovine pancreatic ribonuclease A

Protein Science ◽

10.1110/ps.0242803 ◽

2003 ◽

Vol 12 (5) ◽

pp. 939-952 ◽

Cited By ~ 8

Author(s):

Margherita Ruoppolo ◽

Stefania Orrù ◽

Fabio Talamo ◽

Johanna Ljung ◽

Annamari Pirneskoski ◽

...

Keyword(s):

Protein Disulfide Isomerase ◽

Ribonuclease A ◽

Folding Pathway ◽

Disulfide Isomerase ◽

Pancreatic Ribonuclease ◽

Pancreatic Ribonuclease A ◽

Protein Disulfide

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Reactivity of glutaredoxins 1, 2 and 3 from Escherichia coli and protein disulfide isomerase towards glutathionyl-mixed disulfides in ribonuclease A

FEBS Letters ◽

10.1016/s0014-5793(98)01698-6 ◽

1999 ◽

Vol 443 (2) ◽

pp. 85-88 ◽

Cited By ~ 24

Author(s):

Johanna Lundström-Ljung ◽

Alexios Vlamis-Gardikas ◽

Fredrik Åslund ◽

Arne Holmgren

Keyword(s):

Escherichia Coli ◽

Protein Disulfide Isomerase ◽

Ribonuclease A ◽

Disulfide Isomerase ◽

Mixed Disulfides ◽

Protein Disulfide

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Functional Relationship between Protein Disulfide Isomerase Family Members during the Oxidative Folding of Human Secretory Proteins

Molecular Biology of the Cell ◽

10.1091/mbc.e10-04-0356 ◽

2010 ◽

Vol 21 (18) ◽

pp. 3093-3105 ◽

Cited By ~ 62

Author(s):

Lori A. Rutkevich ◽

Myrna F. Cohen-Doyle ◽

Ulf Brockmeier ◽

David B. Williams

Keyword(s):

Protein Disulfide Isomerase ◽

High Efficiency ◽

Family Members ◽

Secretory Proteins ◽

Human Hepatoma Cells ◽

Oxidative Folding ◽

Disulfide Isomerase ◽

The Impact ◽

Protein Disulfide ◽

Protein Disulfide Isomerase Family

To examine the relationship between protein disulfide isomerase family members within the mammalian endoplasmic reticulum, PDI, ERp57, ERp72, and P5 were depleted with high efficiency in human hepatoma cells, either singly or in combination. The impact was assessed on the oxidative folding of several well-characterized secretory proteins. We show that PDI plays a predominant role in oxidative folding because its depletion delayed disulfide formation in all secretory proteins tested. However, the phenotype was surprisingly modest suggesting that other family members are able to compensate for PDI depletion, albeit with reduced efficacy. ERp57 also exhibited broad specificity, overlapping with that of PDI, but with preference for glycosylated substrates. Depletion of both PDI and ERp57 revealed that some substrates require both enzymes for optimal folding and, furthermore, led to generalized protein misfolding, impaired export from the ER, and degradation. In contrast, depletion of ERp72 or P5, either alone or in combination with PDI or ERp57 had minimal impact, revealing a narrow substrate specificity for ERp72 and no detectable role for P5 in oxidative protein folding.

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