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Key steps in this transformation include ATP binding, closure of a mobile structure (lid) over the nucleotide, and a subsequent 90o rotation of the NTD relative to the MD (Krukenberg et al., 2011).
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Upon agonist binding, lobe closure of the ligand-binding domain produced outward pulling of the M3-D2 leading, leading to outward movements of the C-termini of the pore-lining M3 helices and opening of the channel.
If PAPS binding and closure at the first subunit stabilize the adjacent cap in either the open or closed position, the affinity of PAPS for the second site will decrease relative to that for the first.
In the case of the Cys20-MDCC-conjugated TM-PaDDAH, the fluorescence increased, consistent with l-citrulline binding causing closure of the L1-loop and introducing interactions of the fluorophore with the protein surface.
Instead, our data indicate that the major allosteric transition triggered by ATP binding is closure of kinesin's nucleotide cleft, accompanied by large-scale distortion of the peptide backbone including the central beta sheet.
Similar experiments using expressed smooth muscle myosin suggested R246 is significant for back door closure after ATP binding to the active site and that E469 is important for ATP hydrolysis by its positioning of a water molecule for nucleophilic attack on the γ-phosphate [ 49].
Two pathways jointly propagate the action of substrate-WW binding to produce closure and rigidification of three PPIase catalytic-site loops.
The conformational changes of NBD2 observed here might partially be due to the Mg-ADP binding and full closure of the degenerate site, but we suggest that the Mg-ADP binding on the NBD2 consensus site plays a more prominent role.
It appears that the various key components of the catalytically competent architecture are brought together as separate events, with the formation of strong hydrogen bonding between active site His195 and substrate early in binding and the closure of the catalytically necessary protein surface loop over the bound substrate as the final event of the binding process.
The coupling of PAPS binding and cap closure is depicted in Figure 7.
ABA binding induces the closure of the ligand entry gate onto the latch, allowing the receptor to dock onto PP2Cs.
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