There are proteins given by Kleiger et al. that contain repeats with variable amino acids more closely matching those usually found in FliH (1DBT contains the repeat GLEEG, for instance). However, 1HJR was chosen because it features two identical glycine repeat segments (from identical subunits) that dimerize, whereas the helix containing the glycine repeat in 1DBT dimerizes with a helix that does not contain a GxxxG. Given that two FliH proteins dimerize to form Trametinib datasheet a heterotrimeric complex with FliI [17], and that many
FliH proteins contain several repeats throughout the protein, it seems likely that, in FliH, dimerization would occur between two helices that both contain glycine repeats, making 1HJR a better model than 1DBT. See Figure 9 for a molecular model of the GxxxG helix-helix dimer in this protein. Figure 9 Glycine repeat-mediated interaction between two helices in E. coli site-specific recombinase. The helix-helix interaction in E. coli site-specific recombinase (PDB ID 1HJR) is shown. (A) A side view of the helices that undergo glycine repeat-facilitated BIBW2992 dimerization. The pink squares represent the atoms of the residues in the glycine repeat segment. (B) An end-on view of the same interaction. (C) A more detailed representation of the interactions of the individual residues in
the glycine repeat, viewed from the side. (D) Detailed representation viewed end-on. (A) and (B) were produced using PyMol [34], while (C) and (D) were produced using TURBO-FRODO [33]. Parts (C) and (D) of Figure 9 suggest that interactions between adjacent glycine residues may have an important role in the dimerization process, as the lack of a bulky side chain in this residue allows a C-H… O hydrogen bond to form between the two
Gly Benzatropine residues. In addition, the closest contacts between residues with side chains appear to be between the x1 position in the first helix and the x2 position of the second twofold symmetry-related helix. In the case of 1HJR, the NE of the Arg residue in position x1 donates a hydrogen bond to the OE1 oxygen atom of the Gln residue in x2 on the opposite helix. Although residues in positions x2 and x3 can also make interactions with the adjacent twofold symmetry-related helix, they do not appear to be as close together in space. Discussion Functional significance of the variability in length of glycine repeats in different FliH proteins Given the large amount of variability in the lengths of the glycine repeat segments in different FliH proteins, it begs the question as to whether helix-helix dimerization or some other property inherent to the GxxxG sequences is functionally important in FliH.