5dn5 | pdb_00005dn5
Structure of a C-terminally truncated glycoside hydrolase domain from Salmonella typhimurium FlgJ
Structural highlights
Function[FLGJ_SALTY] Flagellum-specific muramidase which hydrolyzes the peptidoglycan layer to assemble the rod structure in the periplasmic space.[1] Publication Abstract from PubMedFlgJ is a glycoside hydrolase (GH) enzyme belonging to the Carbohydrate Active enZyme (CAZy) family GH73. It facilitates passage of the bacterial flagellum through the peptidoglycan (PG) layer by cleaving the beta-1,4 glycosidic bond between N-acetylglucosamine and N-acetylmuramic acid sugars that comprise the glycan strands of PG. Here we describe the crystal structure of the GH domain of FlgJ from bacterial pathogen Salmonella typhimurium (StFlgJ). Interestingly, the active site of StFlgJ was blocked by the C-terminal alpha-helix of a neighbouring symmetry mate and a beta-hairpin containing the putative catalytic glutamic acid residue Glu223 was poorly resolved and could not be completely modeled into the electron density, suggesting it is flexible. Previous reports have shown that the GH73 enzyme Auto from Listeria monocytogenes is inhibited by an N-terminal alpha-helix that may occlude the active site in similar fashion. To investigate if the C-terminus of StFlgJ inhibits GH activity, the glycolytic activity of StFlgJ was assessed with and without the C-terminal alpha-helix. The GH activity of StFlgJ was unaffected by the presence or absence of the alpha-helix, suggesting it is not involved in regulating activity. Removal of the C-terminal alpha-helix did, however, allow a crystal structure of the domain to be obtained where the flexible beta-hairpin containing residue Glu223 was entirely resolved. The beta-hairpin was positioned such that the active site groove was fully solvent-exposed, placing Glu223 nearly 21.6 A away from the putative general acid/base residue Glu184, which is too far apart for these two residues to coordinate glycosidic bond hydrolysis. The mobile nature of the StFlgJ beta-hairpin is consistent with structural studies of related GH73 enzymes, suggesting that a dynamic active site may be common to many GH73 enzymes, in which the active site opens to capture substrate and then closes to correctly orient active site residues for catalysis. Structural and Biochemical Insights into the Peptidoglycan Hydrolase Domain of FlgJ from Salmonella typhimurium.,Zaloba P, Bailey-Elkin BA, Derksen M, Mark BL PLoS One. 2016 Feb 12;11(2):e0149204. doi: 10.1371/journal.pone.0149204., eCollection 2016. PMID:26871950[2] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
| ||||||||||||||||||||||
