Journal: Nature / Year: 2010 Title: Coupled chaperone action in folding and assembly of hexadecameric Rubisco. Authors: Cuimin Liu / Anna L Young / Amanda Starling-Windhof / Andreas Bracher / Sandra Saschenbrecker / Bharathi Vasudeva Rao / Karnam Vasudeva Rao / Otto Berninghausen / Thorsten Mielke / F Ulrich ...Authors: Cuimin Liu / Anna L Young / Amanda Starling-Windhof / Andreas Bracher / Sandra Saschenbrecker / Bharathi Vasudeva Rao / Karnam Vasudeva Rao / Otto Berninghausen / Thorsten Mielke / F Ulrich Hartl / Roland Beckmann / Manajit Hayer-Hartl / Abstract: Form I Rubisco (ribulose 1,5-bisphosphate carboxylase/oxygenase), a complex of eight large (RbcL) and eight small (RbcS) subunits, catalyses the fixation of atmospheric CO(2) in photosynthesis. The ...Form I Rubisco (ribulose 1,5-bisphosphate carboxylase/oxygenase), a complex of eight large (RbcL) and eight small (RbcS) subunits, catalyses the fixation of atmospheric CO(2) in photosynthesis. The limited catalytic efficiency of Rubisco has sparked extensive efforts to re-engineer the enzyme with the goal of enhancing agricultural productivity. To facilitate such efforts we analysed the formation of cyanobacterial form I Rubisco by in vitro reconstitution and cryo-electron microscopy. We show that RbcL subunit folding by the GroEL/GroES chaperonin is tightly coupled with assembly mediated by the chaperone RbcX(2). RbcL monomers remain partially unstable and retain high affinity for GroEL until captured by RbcX(2). As revealed by the structure of a RbcL(8)-(RbcX(2))(8) assembly intermediate, RbcX(2) acts as a molecular staple in stabilizing the RbcL subunits as dimers and facilitates RbcL(8) core assembly. Finally, addition of RbcS results in RbcX(2) release and holoenzyme formation. Specific assembly chaperones may be required more generally in the formation of complex oligomeric structures when folding is closely coupled to assembly.
History
Deposition
Oct 20, 2009
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Header (metadata) release
Oct 27, 2009
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Map release
Jan 20, 2010
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Update
Jan 20, 2010
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Current status
Jan 20, 2010
Processing site: PDBe / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Details: Quantifoil grids (3/3) with 2 nm carbon on top
Vitrification
Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 85 K / Instrument: OTHER / Details: Vitrification instrument: Vitrobot Method: 10 second blotting before plunging, used 2 layers of filter paper
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Electron microscopy
Microscope
FEI TECNAI 12
Temperature
Average: 95 K
Image recording
Category: CCD / Film or detector model: FEI EAGLE (2k x 2k) / Digitization - Sampling interval: 3.308 µm / Average electron dose: 20 e/Å2 / Details: Data collected on CCD / Bits/pixel: 16
Tilt angle max
0
Electron beam
Acceleration voltage: 120 kV / Electron source: LAB6
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