@article {Srinivasachari2022.07.06.499027, author = {Sathya Srinivasachari and Vikas R. Tiwari and Tripti R. Kharbanda and Ramanathan Sowdamini and Ramaswamy Subramanian}, title = {A Dimer between Monomers and Hexamers {\textendash} Oligomeric Variations in Glucosamine 6-Phosphate Deaminase Family}, elocation-id = {2022.07.06.499027}, year = {2022}, doi = {10.1101/2022.07.06.499027}, publisher = {Cold Spring Harbor Laboratory}, abstract = {In bacteria that live in hosts whose terminal sugar is a sialic acid, Glucosamine 6-phosphate deaminase (NagB) catalyzes the last step in the conversion of sialic acid into Fructose-6-phosphate, which enters the glycolytic pathway. The enzyme exists as a hexamer in Gram-negative bacteria and is shown to be allosterically regulated. In Gram-positive bacteria, it exists as a monomer and lacks allosteric regulation. Our identification of a dimeric Gram-negative bacterial NagB motivated us to characterize the structural basis of the various oligomeric forms. We characterized the crystal structures of NagB from two Gram-negative pathogens, Haemophilus influenzae (Hi) and Pasturella multocida (Pm). The Hi-NagB is active as a hexamer, while Pm-NagB is active as a dimer. We confirm that this is not a crystallographic artifact by cryo-electron microscopy. Both Hi-NagB and Pm-NagB contain the C-terminal helix, and the residues in the interface involved in oligomerization are conserved. The hexamer is described as a dimer of trimers. In the Pm-NagB dimer, the dimeric interface is conserved. This would suggest that the three possible oligomeric forms of NagB are a monomer, a dimer, and a trimer of dimers. Computational modeling and MD simulations indicate that the residues at the trimeric interface have less stabilizing energy of oligomer formation than those in the dimer interface. We propose that Pm-NagB is the evolutionary link between the monomer and the hexamer forms.Competing Interest StatementThe authors have declared no competing interest.}, URL = {https://www.biorxiv.org/content/early/2022/07/06/2022.07.06.499027}, eprint = {https://www.biorxiv.org/content/early/2022/07/06/2022.07.06.499027.full.pdf}, journal = {bioRxiv} }