Thioredoxin Domain Non-equivalence and Anti-chaperone Activity of Protein Disulfide Isomerase Mutants in Vivo

Whiteley, Erik M.; Hsu, Tsu‐An; Betenbaugh, Michael J.

doi:10.1074/jbc.272.36.22556

Cited by 26 publications

(16 citation statements)

References 45 publications

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“…In the latter study, a synergistic effect of co-expression of BiP with PDI was reported, consistent with a previous study from this group reporting the overexpression of PDI alone in yeast significantly improves heterologous protein secretion (Robinson et al, 1994). In insect cells, overexpression of BiP, PDI, or the cytosolic chaperone HSP70 has been shown to improve the solubility and secretion of Mab from baculovirus-infected cells (Ailor and Betenbaugh, 1998;Hsu and Betenbaugh, 1997;Hsu et al, 1996;Whiteley et al, 1997b). In general, it is difficult to draw firm conclusions from these conflicting reports, other than to note that production of mammalian recombinant proteins in yeast or insect cells can be enhanced by the co-expression of mammalian folding chaperones, foldases and their cofactors.…”

Section: Engineering the Rate Of Mab Folding And Assembly Reactionssupporting

confidence: 76%

Engineering mammalian cell factories for improved recombinant monoclonal antibody production: lessons from nature?

Dinnis

James

2005

Biotech & Bioengineering

143

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In this review we consider how cell specific recombinant monoclonal antibody (Mab) production by engineered mammalian cells can be improved. Whilst it is generally recognized that Mab production is limited post-transcriptionally at folding and assembly reactions, genetic engineering strategies based on overexpression of individual chaperones or foldases in mammalian cells have not reliably increased cell specific Mab production. Given that recent studies have established that chaperones and foldases themselves exist in a large multiprotein complex, which may coordinate the sequential processing of Mabs, we propose that global expansion of all components of the secretory pathway will likely be necessary to generically improve recombinant Mab production by mammalian cells. In this context, what can be learnt from nature? Important recent studies have delineated some of the main cellular pathways involved in the differentiation of B-cells into nature's own high level Mab producers, plasma cells. This is achieved by a dramatic re-programming of cellular function where the coordinated expansion of metabolic and secretory machinery precedes Ig production, then is maintained by induction of a key intracellular signaling pathway, the unfolded protein response (UPR). Here we review genetic engineering strategies to increase cell specific production rate and discuss whether manipulation of intracellular signaling systems such as the UPR will provide a novel means to engineer mammalian cells for high level recombinant Mab production.

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Section: Engineering the Rate Of Mab Folding And Assembly Reactionssupporting

confidence: 76%

Engineering mammalian cell factories for improved recombinant monoclonal antibody production: lessons from nature?

Dinnis

James

2005

Biotech & Bioengineering

143

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“…For PDI, the extent to which the chaperone and redox\isomerase activities are apparent in the folding of proteins is substrate-dependent, as is the degree of involvement of individual a or ah domains in redox\isomerase activity [69,79,[82][83][84]. Whereas chaperone activity has been shown for disulphide-containing proteins, such as lysozyme [85,86], and independence of the isomerase and chaperone activities has been shown in itro in the refolding of acidic phospholipase A # , glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and rhodanese [87][88][89], chaperone activity is lacking and does not seem to be required for folding of substrates such as antibody Fab fragments with intact disulphide bonds [90].…”

Section: Chaperone Functionmentioning

confidence: 99%

The protein disulphide-isomerase family: unravelling a string of folds

Ferrari

Söling

1999

Biochemical Journal

406

359

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The mammalian protein disulphide-isomerase (PDI) family encompasses several highly divergent proteins that are involved in the processing and maturation of secretory proteins in the endoplasmic reticulum. These proteins are characterized by the presence of one or more domains of roughly 95-110 amino acids related to the cytoplasmic protein thioredoxin. All but the PDI-D subfamily are composed entirely of repeats of such domains, with at least one domain containing and one domain lacking a redox-active -Cys-Xaa-Xaa-Cys-tetrapeptide. In addition to their known roles as redox catalysts and isomerases, the last few years have revealed additional functions of the PDI proteins,

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“…The mechanism for reduced BPTI secretion upon PDI overexpression is not clear, particularly given the beneficial effects of PDI overexpression for platelet-derived growth factor, Schizosaccharomyces pombe acid phosphatase, and antistasin (20,29). 2 One possible explanation is the "antichaperone" activity of PDI (30), which has also been invoked to explain negative effects of mutant PDI expression on antibody secretion in insect cells (31).…”

Section: Increased Pdi Levels Decrease Secretion Of All Mutants Propomentioning

confidence: 99%

Protein Folding Stability Can Determine the Efficiency of Escape from Endoplasmic Reticulum Quality Control

Kowalski

Parekh

Murai

et al. 1998

Journal of Biological Chemistry

115

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A fraction of each secreted protein is retained and degraded by the endoplasmic reticulum (ER) quality control apparatus that restricts export to correctly folded proteins. The intrinsic biophysical attributes that determine efficiency of escape from this proofreading process have been examined by expressing mutants of bovine pancreatic trypsin inhibitor (BPTI) in yeast. Secretion efficiency is strongly correlated with thermodynamic stability for a series of six point mutations of BPTI. No correlation of secretion efficiency with either oxidative folding or refolding rates in vitro is found; both the rapidly folded Y35L BPTI mutant and the slowly unfolded G36D BPTI mutant exhibit low secretion efficiency. Elimination of cysteines 14 and 38 by mutagenesis does not increase secretion efficiency, indicating that intramolecular thiol/disulfide rearrangements are not primarily responsible for retention and degradation of destabilized BPTI variants. Mutant yeast strains with diminished ER-associated degradation do not secrete BPTI more efficiently, indicating that retention and degradation are separable processes. These data support a model for ER quality control, wherein protein folding is functionally reversible and the relative rates of folding, unfolding, vesicular export, and retention determine secretion efficiency.

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Thioredoxin Domain Non-equivalence and Anti-chaperone Activity of Protein Disulfide Isomerase Mutants in Vivo

Cited by 26 publications

References 45 publications

Engineering mammalian cell factories for improved recombinant monoclonal antibody production: lessons from nature?

Engineering mammalian cell factories for improved recombinant monoclonal antibody production: lessons from nature?

The protein disulphide-isomerase family: unravelling a string of folds

Protein Folding Stability Can Determine the Efficiency of Escape from Endoplasmic Reticulum Quality Control

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