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Introducing a bulkier side chain at Asp778 appears to close the off-pathway cavity from the main channeling path.
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Elongated bubble, the main mechanism of anti-water coning by injecting nitrogen, can block high-permeable water channeling path to a certain extent.
The results show that there are three forms of nitrogen in the water channeling path: tiny bubble, elongated bubble, and gas channeling, which are related to water saturation and gas injection volume in the water channel.
These results were consistent with the predicted function of the crevice as a channeling path.
Introducing residues with bulkier side chains into a predicted channeling path is a useful approach for validating substrate channeling and exploring the structural architecture of an interconnecting path between active sites.
Here, we substituted four residues at different points along the predicted channeling path in BjPutA with bulkier side chains.
Structural analysis of the channeling path in BjPutA provides new insight into how P5C/GSA is shuttled between the PRODH and P5CDH active sites.
Replacing Asp779 with Tyr decreased the internal width of the predicted channeling path between helices 770s (residues 773 785) and α5a by 2.5 Å or ∼25%.
PutA also undergoes a conformational change upon flavin reduction, with a conserved ion pair (Arg456 Glu197) proposed to act as a gate between the PRODH domain and the main channeling pathway.
The main combustion path goes via ethylene.
Hydraulic fracture is the main flow path for gas transport.
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