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dc.contributor.authorStuart, SF
dc.contributor.authorLeatherbarrow, RJ
dc.contributor.authorWillison, KR
dc.date.accessioned2018-08-17T09:15:11Z
dc.date.issued2011-01-07
dc.identifier1
dc.identifier.citationJOURNAL OF BIOLOGICAL CHEMISTRY, 2011, 286 pp. 178 - 184
dc.identifier.issn0021-9258
dc.identifier.urihttps://repository.icr.ac.uk/handle/internal/2342
dc.identifier.doi10.1074/jbc.M110.166256
dc.description.abstractActin requires the chaperonin containing TCP1 (CCT), a hexadecameric ATPase essential for cell viability in eukaryotes, to fold to its native state. Following binding of unfolded actin to CCT, the cavity of the chaperone closes and actin is folded and released in an ATP-dependent folding cycle. In yeast, CCT forms a ternary complex with the phosducin-like protein PLP2p to fold actin, and together they can return nascent or chemically denatured actin to its native state in a pure in vitro folding assay. The complexity of the CCT-actin system makes the study of the actin folding mechanism technically challenging. We have established a novel spectroscopic assay through selectively labeling the C terminus of yeast actin with acrylodan and observe significant changes in the acrylodan fluorescence emission spectrum as actin is chemically unfolded and then refolded by the chaperonin. The variation in the polarity of the environment surrounding the fluorescent probe during the unfolding/folding processes has allowed us to monitor actin as it folds on CCT. The rate of actin folding at a range of temperatures and ATP concentrations has been determined for both wild type CCT and a mutant CCT, CCT4anc2, defective in folding actin in vivo. Binding of the non-hydrolysable ATP analog adenosine 5’-(beta,gamma-imino)-triphosphate to the ternary complex leads to 3-fold faster release of actin from CCT following addition of ATP, suggesting a two-step folding process with a conformational change occurring upon closure of the cavity and a subsequent final folding step involving packing of the C terminus to the native-like state.
dc.format.extent178 - 184
dc.languageeng
dc.language.isoeng
dc.publisherAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
dc.titleA Two-step Mechanism for the Folding of Actin by the Yeast Cytosolic Chaperonin
dc.typeJournal Article
rioxxterms.versionofrecord10.1074/jbc.M110.166256
rioxxterms.licenseref.startdate2011-01-07
rioxxterms.typeJournal Article/Review
dc.relation.isPartOfJOURNAL OF BIOLOGICAL CHEMISTRY
pubs.notesaffiliation: Willison, KR (Reprint Author), 237 Fulham Rd, London SW3 6JB, England. Stuart, Sarah F.; Leatherbarrow, Robin J., Univ London Imperial Coll Sci Technol & Med, Inst Biol Chem, London SW7 2AZ, England. Stuart, Sarah F.; Willison, Keith R., Inst Canc Res, Sect Cell & Mol Biol, London SW3 6JB, England. keywords-plus: EUKARYOTIC CHAPERONIN; CCT; INTERACTS; PROTEIN; STATE research-areas: Biochemistry & Molecular Biology web-of-science-categories: Biochemistry & Molecular Biology author-email: [email protected] researcherid-numbers: Leatherbarrow, Robin/H-3070-2014 orcid-numbers: Leatherbarrow, Robin/0000-0001-8193-5964 funding-acknowledgement: Engineering and Physical Sciences Research Council through the Institute of Chemical Biology funding-text: Supported by the Engineering and Physical Sciences Research Council through the Institute of Chemical Biology. number-of-cited-references: 24 times-cited: 14 usage-count-last-180-days: 0 usage-count-since-2013: 10 journal-iso: J. Biol. Chem. doc-delivery-number: 700YZ unique-id: ISI:000285782800019 oa: gold_or_bronze da: 2018-08-16
pubs.notesNot known
pubs.organisational-group/ICR
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Closed research teams
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Closed research teams/Chromatin Regulation
pubs.organisational-group/ICR
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Closed research teams
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Closed research teams/Chromatin Regulation
pubs.volume286
pubs.embargo.termsNot known
icr.researchteamChromatin Regulationen_US
dc.contributor.icrauthorWillison, Keithen


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