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Your search keyword '"Group I Chaperonins genetics"' showing total 3 results
Start Over Descriptor "Group I Chaperonins genetics" Topic ribulose-bisphosphate carboxylase
3 results on '"Group I Chaperonins genetics"'

1. Improved recombinant expression and purification of functional plant Rubisco.

Author

Wilson RH, Thieulin-Pardo G, Hartl FU, and Hayer-Hartl M

Subjects
Arabidopsis genetics, Arabidopsis Proteins metabolism, Carbon Dioxide metabolism, Escherichia coli genetics, Escherichia coli metabolism, Gene Expression, Genetic Vectors chemistry, Genetic Vectors metabolism, Group I Chaperonins metabolism, Histidine genetics, Histidine metabolism, Kinetics, Models, Molecular, Molecular Chaperones metabolism, Oligopeptides genetics, Oligopeptides metabolism, Phosphate-Binding Proteins metabolism, Photosynthesis genetics, Protein Folding, Protein Isoforms genetics, Protein Isoforms metabolism, Protein Structure, Secondary, Protein Subunits genetics, Protein Subunits isolation & purification, Protein Subunits metabolism, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins isolation & purification, Recombinant Fusion Proteins metabolism, Ribulose-Bisphosphate Carboxylase isolation & purification, Ribulose-Bisphosphate Carboxylase metabolism, Arabidopsis enzymology, Arabidopsis Proteins genetics, Cloning, Molecular methods, Group I Chaperonins genetics, Molecular Chaperones genetics, Phosphate-Binding Proteins genetics, Ribulose-Bisphosphate Carboxylase genetics
Abstract

Improving the performance of the key photosynthetic enzyme Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) by protein engineering is a critical strategy for increasing crop yields. The extensive chaperone requirement of plant Rubisco for folding and assembly has long been an impediment to this goal. Production of plant Rubisco in Escherichia coli requires the coexpression of the chloroplast chaperonin and four assembly factors. Here, we demonstrate that simultaneous expression of Rubisco and chaperones from a T7 promotor produces high levels of functional enzyme. Expressing the small subunit of Rubisco with a C-terminal hexahistidine-tag further improved assembly, resulting in a ~ 12-fold higher yield than the previously published procedure. The expression system described here provides a platform for the efficient production and engineering of plant Rubisco., (© 2019 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

Published
2019
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2. Plant RuBisCo assembly in E. coli with five chloroplast chaperones including BSD2.

Author

Aigner H, Wilson RH, Bracher A, Calisse L, Bhat JY, Hartl FU, and Hayer-Hartl M

Subjects
Arabidopsis Proteins chemistry, Arabidopsis Proteins genetics, Chaperonin 60 chemistry, Chaperonin 60 genetics, Chaperonin 60 metabolism, Chloroplasts metabolism, Crystallography, X-Ray, Group I Chaperonins chemistry, Group I Chaperonins genetics, Group I Chaperonins metabolism, Molecular Chaperones chemistry, Molecular Chaperones genetics, Mutagenesis, Protein Folding, Recombinant Proteins chemistry, Recombinant Proteins genetics, Ribulose-Bisphosphate Carboxylase chemistry, Ribulose-Bisphosphate Carboxylase genetics, Arabidopsis enzymology, Arabidopsis Proteins metabolism, Escherichia coli enzymology, Molecular Chaperones metabolism, Recombinant Proteins metabolism, Ribulose-Bisphosphate Carboxylase metabolism
Abstract

Plant RuBisCo, a complex of eight large and eight small subunits, catalyzes the fixation of CO 2 in photosynthesis. The low catalytic efficiency of RuBisCo provides strong motivation to reengineer the enzyme with the goal of increasing crop yields. However, genetic manipulation has been hampered by the failure to express plant RuBisCo in a bacterial host. We achieved the functional expression of Arabidopsis thaliana RuBisCo in Escherichia coli by coexpressing multiple chloroplast chaperones. These include the chaperonins Cpn60/Cpn20, RuBisCo accumulation factors 1 and 2, RbcX, and bundle-sheath defective-2 (BSD2). Our structural and functional analysis revealed the role of BSD2 in stabilizing an end-state assembly intermediate of eight RuBisCo large subunits until the small subunits become available. The ability to produce plant RuBisCo recombinantly will facilitate efforts to improve the enzyme through mutagenesis., (Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

Published
2017
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3. Asymmetric functional interaction between chaperonin and its plastidic cofactors.

Author

Guo P, Jiang S, Bai C, Zhang W, Zhao Q, and Liu C

Subjects
Algal Proteins agonists, Algal Proteins chemistry, Algal Proteins genetics, Arabidopsis metabolism, Arabidopsis Proteins agonists, Arabidopsis Proteins chemistry, Arabidopsis Proteins genetics, Bacterial Proteins agonists, Bacterial Proteins chemistry, Bacterial Proteins genetics, Chaperonin 10 agonists, Chaperonin 10 chemistry, Chaperonin 10 genetics, Chaperonin 10 metabolism, Chaperonin 60 agonists, Chaperonin 60 chemistry, Chaperonin 60 genetics, Chaperonin 60 metabolism, Chlamydomonas metabolism, Escherichia coli metabolism, Escherichia coli Proteins agonists, Escherichia coli Proteins chemistry, Escherichia coli Proteins genetics, Escherichia coli Proteins metabolism, Group I Chaperonins agonists, Group I Chaperonins chemistry, Group I Chaperonins genetics, Molecular Weight, Protein Multimerization, Protein Refolding, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins metabolism, Recombinant Proteins chemistry, Recombinant Proteins metabolism, Rhodospirillum rubrum enzymology, Rhodospirillum rubrum metabolism, Ribulose-Bisphosphate Carboxylase chemistry, Ribulose-Bisphosphate Carboxylase genetics, Algal Proteins metabolism, Arabidopsis Proteins metabolism, Bacterial Proteins metabolism, Chloroplasts metabolism, Group I Chaperonins metabolism, Models, Molecular, Ribulose-Bisphosphate Carboxylase metabolism
Abstract

The specific cochaperonin, chloroplast chaperonin (Cpn)20, consisting of two tandem GroES-like domains, is present abundantly in plant and algal chloroplasts, in addition to Cpn10, which is similar in size to GroES. How Cpn20 oligomers, containing six or eight 10-kDa domains, cooperate with the heptameric ring of chaperonin at the same time as encountering symmetry mismatch is unclear. In the present study, we characterized the functional cooperation of cochaperonins, including two plastidic Cpn20 homo-oligomers from Arabidopsis (AtCpn20) and Chlamydomonas (CrCPN20), and one algal CrCPNs hetero-oligomer, consisting of three cochaperonins, CrCPN11, CrCPN20 and CrCPN23, with two chaperonins, Escherichia coli GroEL and Chlamydomonas CrCPN60. AtCpn20 and CrCPNs were functional for assisting both chaperonins in folding model substrates ribulose bisphosphate carboxylase oxygenase from Rhodospirillum rubrum (RrRubisco) in vitro and complementing GroES function in E. coli. CrCPN20 cooperated only with CrCPN60 (and not GroEL) to refold RrRubisco in vitro and showed differential complementation with the two chaperonins in E. coli. Cochaperonin concatamers, consisting of six to eight covalently linked 10-kDa domains, were functionally similar to their respective native forms. Our results indicate that symmetrical match between chaperonin and cochaperonin is not an absolute requisite for functional cooperation., (© 2015 FEBS.)

Published
2015
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