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For example, Arg53 in the haem-binding pocket of Mtb SeMet-BfrA has flipped orientation compared to that of Arg53 in M. smegmatis Bfr (Figure 5).
Comparison of the ferroxidase centre of Mtb BfrB with that of E. coli ferritin reveals the subtle differences: slightly altered conformations of E131 and E134 as compared to E126 and E129 respectively of E. coli, and flipped orientation of E54 in Mtb BfrB that is stabilized by formation of a salt bridge with R180.
As noted above, the methoxy group is also too large for this active site volume in the alternative, "flipped" orientation.
Plasmids pDD1262 (flipped orientation of the tDNA) and pDD1272 (tdnaΔ:: ETC9) were created by cloning EcoRI– BamHI-digested PCR-amplified fragments using DDO1468/-1469 (flip), or DDO1534/-1535 (ETC9), respectintoy, into EcoRI– BamHI-digested pDD1260.
The alternative, "flipped" orientation is highly disfavored in wild-type OYE 2.6 because the methoxy moiety is too large to fit in the pocket partially defined by Tyr78 and Ile113.
Interestingly, we found that HBA could be fitted best into the electron density in two orientations, depending on the molecule: a first orientation with its aldehyde moiety in hydrogen bonding distance to amino acids H191/N194 and a second, flipped orientation similar to that seen for wild type OYE1.
Similar(44)
Using the criteria described above, we modeled reasonable complexes of alkene 3 in both the "normal" and "flipped" orientations with wild-type OYE 2.6 as well as the Tyr78Trp and Tyr78Trp/Ile113Cys mutants.
In order to flip orientation, use the "-y" option at startup, or keyboard "y" interactively.
In particular, it is unlikely that P5C or GSA can flip orientation while in the tunnel, and torsional motion of GSA is probably restricted.
To prepare the structure for docking, we deleted all bound ligands and crystallographic waters and flipped the orientation of residue Asn 294 in the binding site (the sidechain amide nitrogen and oxygen had been assigned incorrectly).
The Arg53 in Mtb BfrA has flipped its orientation by formation of a salt bridge with Glu57, which in M. smegmatis is mutated to Thr57.
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