What question did this study set out to answer?

To evaluate how controlled coexpression of DnaK-DnaJ-GrpE and GroEL-GroES chaperone systems influences the folding, stability, and aggregation of the recombinant cedar pollen allergen Cryj2 in Escherichia coli.

May 1, 1998Applied and Environmental MicrobiologyOpen Access

Chaperone Coexpression Plasmids: Differential and Synergistic Roles of DnaK-DnaJ-GrpE and GroEL-GroES in Assisting Folding of an Allergen of Japanese Cedar Pollen, Cryj2, in Escherichia coli

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Authors

KNKazuyo NishiharaNara National Research Institute for Cultural Properties MKMasaaki KanemoriKanazawa University MKMasanari KitagawaTakara (Japan)

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Implication

Experimental study demonstrates chaperone coexpression enhances Cryj2 allergen folding in Escherichia coli, indicating synergistic roles in preventing recombinant protein aggregation.

Key Points

To evaluate how controlled coexpression of DnaK-DnaJ-GrpE and GroEL-GroES chaperone systems influences the folding, stability, and aggregation of the recombinant cedar pollen allergen Cryj2 in Escherichia coli.
Constructed a dual-promoter pACYC184-based plasmid permitting independent, dose-dependent induction of DnaK-DnaJ-GrpE via L-arabinose and GroEL-GroES via tetracycline.
Assayed Cryj2 stability, accumulation, and aggregation across wild-type E. coli K-12 and defined mutant strains lacking specific chaperone subunits or the heat-shock transcription factor sigma 32.
Controlled coexpression of DnaK-DnaJ-GrpE, GroEL-GroES, or both systems markedly enhanced Cryj2 stability and yield without extensive aggregation.
Chaperone-deficient and sigma 32-deficient mutants revealed distinct functional folding mechanisms for each chaperone team, along with temperature-dependent synergistic effects in preventing aggregation.

Cite This Study

Nishihara et al. (1998) studied this question.

synapsesocial.com/papers/6a955cd38ce716a7e9fa71c3 https://doi.org/10.1128/aem.64.5.1694-1699.1998

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Implication

Experimental study demonstrates chaperone coexpression enhances Cryj2 allergen folding in Escherichia coli, indicating synergistic roles in preventing recombinant protein aggregation.

Chaperone Coexpression Plasmids: Differential and Synergistic Roles of DnaK-DnaJ-GrpE and GroEL-GroES in Assisting Folding of an Allergen of Japanese Cedar Pollen, Cryj2, in Escherichia coli

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Key Points

Cite This Study

Chaperone Coexpression Plasmids: Differential and Synergistic Roles of DnaK-DnaJ-GrpE and GroEL-GroES in Assisting Folding of an Allergen of Japanese Cedar Pollen, Cryj2, in Escherichia coli

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