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In this study, we used NMR titration and STD to probe the ligand protein binding interface as it exists in solution and combined information from those experiments with bound ligand conformation information from trNOE experiments.
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Importantly, this structure revealed a more extended and distinct binding interface, as well as an altered Nrf2 peptide conformation consisting of an N-terminal helix (Leu19 Arg25) and two short 310 helices (Ile28 to Leu30 and Arg34 to Phe37).
At the same time, strains in different genogroups that use different binding interfaces, as defined by their locations and sequence motifs, can recognize the same HBGA-targets, pointing to the overall functional and structural similarity of these distinct binding sites.
Because the single intrinsic W325 is located on the finger subdomain and is far from the binding interface, it was used as a control site.
Relatedly, how easily does orthologous neofunctionalization occur and how dependent upon protein fold and binding interface size is it?
This β-Gal binding site is conserved among all BV, NV, FUV and TCH with known crystal structures (Bu et al., 2008; Choi et al., 2008; Kubota et al., 2012; Shanker et al., 2014) and thus may be defined as the central binding pocket (CBP) of GI HBGA binding interfaces, because it plays the central role in GI NoV-HBGA interactions, as proposed previously (Tan and Jiang, 2014).
Glu379 is located in domain 3b, structurally distant from any of the predicted syntaxin binding interfaces, and as such it is not surprising that mutational insertion here had no effect on measured closed-conformation syntaxin binding.
P268 constitutes part of the nucleotide-binding interface where it interacts with the adenine moiety of ADP.
Y179 is located on the DNA-binding interface, where it recognizes 8-oxoG A mispairings and stabilizes protein–DNA complexes.
This ability is derived from the strength of molecular binding to the membrane interface as well as from the cohesiveness of the membrane itself.
A highly conserved fucose (Fuc -binding pocket at the center oFuc -bindingg interface is identified as central binding pocket (CBP) that plays a major role in intheacenterwith HBGAs via the α-1,2 Fuc of α-1,3/4 Fuc as the major binding saccharinterfaceSs) (Tan and Jisng, 2014).
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