Uncovering a novel molecular mechanism for scavenging sialic acids in bacteria.

The Journal of Biological Chemistry
Andrew BellNathalie Juge

Abstract

The human gut symbiont Ruminococcus gnavus scavenges host-derived N-acetylneuraminic acid (Neu5Ac) from mucins by converting it to 2,7-anhydro-Neu5Ac. We previously showed that 2,7-anhydro-Neu5Ac is transported into R. gnavus ATCC 29149 before being converted back to Neu5Ac for further metabolic processing. However, the molecular mechanism leading to the conversion of 2,7-anhydro-Neu5Ac to Neu5Ac remained elusive. Using 1D and 2D NMR, we elucidated the multistep enzymatic mechanism of the oxidoreductase (RgNanOx) that leads to the reversible conversion of 2,7-anhydro-Neu5Ac to Neu5Ac through formation of a 4-keto-2-deoxy-2,3-dehydro-N-acetylneuraminic acid intermediate and NAD+ regeneration. The crystal structure of RgNanOx in complex with the NAD+ cofactor showed a protein dimer with a Rossman fold. Guided by the RgNanOx structure, we identified catalytic residues by site-directed mutagenesis. Bioinformatics analyses revealed the presence of RgNanOx homologues across Gram-negative and Gram-positive bacterial species and co-occurrence with sialic acid transporters. We showed by electrospray ionization spray MS that the Escherichia coli homologue YjhC displayed activity against 2,7-anhydro-Neu5Ac and that E. coli could cataboliz...Continue Reading

References

Oct 5, 1990·Journal of Molecular Biology·S F AltschulD J Lipman
Feb 14, 2002·Structure·Simon T M AllardJames H Naismith
Mar 10, 2004·Microbiology and Molecular Biology Reviews : MMBR·Eric R VimrSusan M Steenbergen
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Paul Emsley, Kevin Cowtan
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·E Krissinel, K Henrick
Sep 24, 2005·Infection and Immunity·Deborah M B PostRobert S Munson
Dec 22, 2005·Journal of the American Chemical Society·Louise L MajorJames H Naismith
Apr 25, 2007·Nucleic Acids Research·Ian W DavisDavid C Richardson
Sep 5, 2007·Microbiology·Emmanuele SeveriGavin H Thomas
Oct 7, 2008·Journal of Molecular Biology·Guogang XuGarry L Taylor
Nov 26, 2008·Chemico-biological Interactions·Bengt PerssonUdo Oppermann
Jan 31, 2009·Proceedings of the National Academy of Sciences of the United States of America·Christopher MulliganGavin H Thomas
May 28, 2009·BMC Evolutionary Biology·Salvador Almagro-Moreno, E Fidelma Boyd
Sep 15, 2009·Journal of Bacteriology·Susan M SteenbergenEric R Vimr
Jul 22, 2010·The American Journal of Gastroenterology·Chin Wen PngTimothy H J Florin
Dec 30, 2010·Infection and Immunity·Carolyn MarionSamantha J King
Apr 5, 2011·Acta Crystallographica. Section D, Biological Crystallography·Garib N MurshudovAlexei A Vagin
Nov 1, 2011·Cell Host & Microbe·Angela MarcobalJustin L Sonnenburg
Aug 13, 2013·Journal of Bacteriology·Kathryn A KalivodaEric R Vimr
Jul 10, 2014·Acta Crystallographica. Section D, Biological Crystallography·Thanuja Gangi SettyS Ramaswamy
Jul 31, 2014·IUCrJ·Robbie P JoostenAnastassis Perrakis
Jan 11, 2016·Protein Science : a Publication of the Protein Society·Helena TabermanJuha Rouvinen
Feb 11, 2016·Biochemical Society Transactions·Nathalie JugeC David Owen
Jun 11, 2016·Biochemical Society Transactions·Gavin H Thomas

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Citations

Apr 6, 2021·PLoS Pathogens·Alexandria J HammondJeffrey N Weiser
Apr 26, 2021·The Journal of Biological Chemistry·Takfarinas KentacheEmile Van Schaftingen
Aug 27, 2021·The Journal of Biological Chemistry·Michael J CurrieRenwick C J Dobson
Sep 7, 2021·Chemistry : a European Journal·Xander SchaapkensTiddo J Mooibroek

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BETA
NMR
PCR
gel filtration
size-exclusion chromatography

Software Mentioned

Refmac
DALI
COOT
BLASTp
Phenix AutoBuild
Molprobity
PDB REDO
BLAST
MrBUMP Sculptor pipeline
REFMAC5

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