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TitleMechanism of Enzyme Repair by the AAA Chaperone Rubisco Activase.
Journal, issue, pagesMol Cell, Vol. 67, Issue 5, Page 744-756.e6, Year 2017
Publish dateSep 7, 2017
AuthorsJavaid Y Bhat / Goran Miličić / Gabriel Thieulin-Pardo / Andreas Bracher / Andrew Maxwell / Susanne Ciniawsky / Oliver Mueller-Cajar / John R Engen / F Ulrich Hartl / Petra Wendler / Manajit Hayer-Hartl /
PubMed AbstractHow AAA+ chaperones conformationally remodel specific target proteins in an ATP-dependent manner is not well understood. Here, we investigated the mechanism of the AAA+ protein Rubisco activase (Rca) ...How AAA+ chaperones conformationally remodel specific target proteins in an ATP-dependent manner is not well understood. Here, we investigated the mechanism of the AAA+ protein Rubisco activase (Rca) in metabolic repair of the photosynthetic enzyme Rubisco, a complex of eight large (RbcL) and eight small (RbcS) subunits containing eight catalytic sites. Rubisco is prone to inhibition by tight-binding sugar phosphates, whose removal is catalyzed by Rca. We engineered a stable Rca hexamer ring and analyzed its functional interaction with Rubisco. Hydrogen/deuterium exchange and chemical crosslinking showed that Rca structurally destabilizes elements of the Rubisco active site with remarkable selectivity. Cryo-electron microscopy revealed that Rca docks onto Rubisco over one active site at a time, positioning the C-terminal strand of RbcL, which stabilizes the catalytic center, for access to the Rca hexamer pore. The pulling force of Rca is fine-tuned to avoid global destabilization and allow for precise enzyme repair.
External linksMol Cell / PubMed:28803776
MethodsEM (single particle)
Resolution3.39 - 7.56 Å
Structure data

3699

5nv3

EMDB-3699, PDB-5nv3:
Structure of Rubisco from Rhodobacter sphaeroides in complex with CABP
Method: EM (single particle) / Resolution: 3.39 Å

EMDB-3700:
Structure of Rubisco from Rhodobacter sphaeroides in complex with CABP
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-3701:
Structure of Rubisco from Rhodobacter sphaeroides in complex with CABP and RcaCC
Method: EM (single particle) / Resolution: 7.56 Å

EMDB-3702:
Structure of Rubisco from Rhodobacter sphaeroides in complex with CABP and RcaCC
Method: EM (single particle) / Resolution: 6.05 Å

Chemicals

ChemComp-CAP:
2-CARBOXYARABINITOL-1,5-DIPHOSPHATE

ChemComp-MG:
Unknown entry

Source
  • rhodobacter sphaeroides (bacteria)
KeywordsLYASE / beta barrel

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