ProfileMark Nitz received his BSc in Chemistry from the University of British Columbia in 1995. After earning his PhD with David Bundle at the University of Alberta in 2001, Professor Nitz worked as a postdoctoral fellow with Barbara Imperiali at the Massachusetts Institute of Technology (2001-2004). He started his independent career in the Chemistry Department at the University of Toronto in 2004 as an Assistant Professor. He was promoted to Associate Professor in 2009 and to Professor, in the same department, in 2014. Professor Nitz’s research program spans the areas of synthetic carbohydrate chemistry and chemical biology. Focusing on bacterial biofilm formation the Nitz lab has efficiently synthesized key carbohydrate adhesin’s that form the structure of the biofilm matrix. These compounds have served as substrates to elucidate the biochemistry and structure of the enzymes required for polysaccharide adhesin synthesis and biofilm formation. In addition to Professor Nitz’s work in the glycosciences his lab was part of the team that has developed reagents for use with Mass Cytometry, a technique that is revolutionizing multiparameter cytometric analysis. New chemistry in the Nitz lab is now exploring small molecule reagents for us with Mass Cytometry that will lead to probes able to report on enzyme activity, metabolic state and transporters. GlycoNet Funded ProjectsProducts
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Synthesis of defined mono-de-N-acetylated beta-(1,6)-N-acetyl-d-glucosamine oligosaccharides to characterize PgaB hydrolase activityMono-de-N-acetylated β-(1→6)-N-acetyl-D-glucosamine penta- and hepta-saccharides were obtained using a convergent synthesis. The site of de-N-acetylation drives the selectivity of hydrolysis by PgaB.AM-2 Hospital for Sick Children, University of Toronto | Publication | 2019-11-21 | | Ega3 from the fungal pathogen Aspergillus fumigatus is an endo-$\upalpha$-1,4-galactosaminidase that disrupts microbialbiofilms University of Toronto, McGill University, Hospital for Sick Children | Publication | 2019-08-01 | Natalie Bamford, François LeMauff, Adithya S Subramanian, Patrick Yip, Claudia Millán, Yongzhen Zhang, Caitlin Zacharias, Adam Forman, Mark Nitz, Jeroen D C Codée, Isabel Usón, Donald Sheppard, Patricia Lynne Howell | NSERC - MorrisonNSERC masters award $17500 per yearAM-2 | Award | 2016-09-01 | Zachary Morrison | Monosaccharide inhibitors targeting carbohydrate esterase family 4 de-N-acetylasesAM-2 University of Toronto | Publication | 2018-10-01 | Benjamin DiFrancesco, Zachary Morrison, Mark Nitz | The unique solution structure and immunochemistry of the Candida albicans b-1,2-mannopyranan cell wall antigens. University of Toronto | Publication | 2002-02-01 | Mark Nitz, Ling C-C, Otter A, Cutler JE, Bundle DR | Horace S. Isabell AwardThe Horace S. Isbell Award acknowledges excellence in and promise of continued quality of contribution to research in carbohydrate chemistry. The winner must be under the age of 45 at the time of the award.AM-2, CD-7, TP-11 University of Toronto | Award | 2016-02-28 | | Densely functionalized $\upbeta$-CDs via solid-supported synthesis University of Toronto | Publication | 2018-02-01 | | Characterization of E. coli PgaB,
an enzyme essential for biofilm formation University of Toronto | Publication | 2011-07-01 | J. Poloczek, A. Chibba, Mark Nitz, D. Little and P.L. Howell | Metal-dependent Polysaccharide Deacetylase PgaB. University of Toronto, Hospital for Sick Children | Publication | 2014-06-01 | | Structural insights for the N-deacetylation of poly-β-1,6-N-acetyl-D-glucosamine University of Toronto, Hospital for Sick Children | Publication | 2012-11-01 | | Structural insights for the Ndeacetylation
of poly-β-1,6-N-acetyl-D-glucosamine University of Toronto, Hospital for Sick Children | Publication | 2011-06-01 | | Structural insights for the N-deacetylation of poly-b-1,6-N-acetyl-d-glucosamine University of Toronto, Hospital for Sick Children | Publication | 2011-02-01 | | Development of mass cytometry methods for bacterial discrimination University of Guelph, University of Toronto | Publication | 2011-12-01 | | Structural and Functional Characterization of the Poly-β-1,6-N-Acetyl-D- Glucosamine (PNAG) Deacetylase IcaB University of Toronto, Hospital for Sick Children | Publication | 2014-06-01 | | Synthesis and evaluation of inhibitors of E. coli PgaB, a polysaccharide de-N-acetylase involved in biofilm formation. Hospital for Sick Children, University of Toronto | Publication | 2012-09-01 | | Combining in situ proteolysis and mass spectrometry to crystallize Escherichia coli PgaB. University of Toronto, Hospital for Sick Children | Publication | 2012-07-01 | | The structure and metal dependent activity of Escherichia coli PgaB provides insight into the partial de-N-acetylation of poly-β-1,6-N-acetyl-D-glucosamine. University of Toronto, Hospital for Sick Children | Publication | 2012-07-01 | | Organotellurium scaffolds for mass cytometry reagent development University of Toronto | Publication | 2015-01-01 | | PgaB is required for the De-N-acetylation and recognition of the biofilmpolysaccharide PNAG University of Toronto, Hospital for Sick Children | Publication | 2013-07-01 | | PgaB is required for the De-N-acetylation and recognition of the biofilm polysaccharide PNAG University of Toronto, Hospital for Sick Children | Publication | 2013-07-01 | | Functional Characterization of Staphylococcus epidermidis IcaB, a De-N-acetylase Important for Biofilm Formation. Hospital for Sick Children, University of Toronto | Publication | 2013-08-01 | | Tellurium-based mass cytometry barcode for live and fixed cells University of Toronto | Publication | 2018-07-01 | | Modification and periplasmic translocation of the biofilm exopolysaccharide Poly-β-1,6-N-acetyl-D-glucosamine. University of Toronto, Hospital for Sick Children | Publication | 2014-07-01 | | Lanthanide nanoparticles for high sensitivity multiparameter single cell analysis University of Toronto | Publication | 2019-01-01 | Jothirmayanantham Pichaandi, Guangyao Zhao, Alexandre Bouzekri, Elsa Lu, Olga Ornatsky, Vladimir Baranov, Mark Nitz, Mitchell A Winnik | TePhe, a tellurium-containing phenylalanine mimic, allows monitoring of protein synthesis in vivo with mass cytometry University of Alberta, University of Toronto | Publication | 2019-04-01 | | De-N-acetylation and export of the Biofilm Exopolysaccharide Poly-β-1,6-N-acetyl-D-glucosamine (PNAG) University of Toronto, Hospital for Sick Children | Publication | 2014-08-01 | | Modification and Translocation of the BiofilmExopolysaccharide Poly-b-1,6-N-acetyl-d-glucosamine (PNAG) University of Toronto, Hospital for Sick Children | Publication | 2014-06-01 | | Deacetylation of galactosaminogalactan in Aspergillus fumigatus is a required post-synthesis modification for adherence and virulence McGIll University, University of Toronto, Hospital for Sick Children | Publication | 2014-04-01 | M.J. Lee, A. Geller, F.N. Gravelat, Hannah Liu, B. D. Snarr, S. D. Baptista, Dao Nguyen, Mark Nitz, Patricia Lynne Howell, S. G. Filler, And D. C. Sheppard | Developing a Small Molecule Inhibitor of the Psuedomonas aeruginosa Biofilm Exopolysaccharide Deacetylase PelAPoster Presentation at ASM Biofilms 2019 conference in Washington, D.C. AM-2 University of Toronto, Hospital for Sick Children | Presentation | 2019-10-11 | | Developing a Small Molecule Inhibitor of the Psuedomonas aeruginosa Biofilm Exopolysaccharide Deacetylase PelALightning talk at GlycoNet 2019 Symposium AM-2 University of Toronto, Hospital for Sick Children | Presentation | 2019-05-15 | | PgaB orthologues contain a glycoside hydrolase domain that cleaves deacetylated poly-$\upbeta$(1,6)-N-acetylglucosamine and can disrupt bacterial biofilmsAM-15 University of Toronto, McGill University, Hospital for Sick Children | Publication | 2018-04-01 | Dustin J Little, Roland Pfoh, François LeMauff, Natalie Bamford, Christina Notte, Perrin Baker, Manita Guragain, Howard Robinson, Gerald B Pier, Mark Nitz, Rajendar Deora, Donald Sheppard, Patricia Lynne Howell | Molecular basis for PNAG-dependent biofilm disruption by PgaBPoly-beta(1,6)-N-acetyl-D-glucosamine (PNAG) is a major biofilm component of many pathogenic bacteria such as Escherichia coli, Bordetella pertussis, Staphylococcus aureus, Staphylococcus epidermidis, and Klebsiella pneumoniae. In Gram-negative bacteria like E. coli and Bordetella spp. PNAG production, processing and export involves the protein products encoded by the pgaABCD operon. PgaB is a two-domain periplasmic protein with a N-terminal domain responsible for de-N-acetylation of PNAG oligomers and a C-terminal PNAG binding domain critical for export. Here, we show that the C-terminal domains of both Bordetella bronchisteptica PgaB (PgaBBb, formerly known as BpsB) and E. coli PgaB (PgaBEc) are functional glycoside hydrolases that cleave de-N-acetylated PNAG (dPNAG) purified from Staphylococcus aureus. We also show that PgaBBb disrupts PNAG-dependent biofilms formed by B. pertussis, S. epidermidis, S. carnosus and E. coli, and potentiates bacterial killing by gentamicin. To gain insight into the relationship between the PgaB de-N-acetylation and glycoside hydrolase functions, we developed an assay using PgaCDEc for the in situ production of PNAG. Mass spectrometry (MS) analysis suggests that de-N-acetylation of this polymer is required prior to its hydrolysis by PgaB. The MS analysis of the PgaB-hydrolyzed dPNAG substrate showed a GlcN-GlcNAc-GlcNAc motif at the new reducing end of detected fragments. Our 1.76 Å crystal structure of the C-terminal domain of PgaBBb reveals a central cavity within an elongated surface groove that appears ideally suited to recognize the GlcN-GlcNAc-GlcNAc motif. The structure, in conjunction with molecular modeling and site directed mutagenesis led to the identification of the dPNAG binding subsites and the probable catalytic acid. Our findings shed light on the role of PgaB within the PNAG production machinery and support a cleavage mechanism that specifically recognizes de-N-acetylated PNAG. Furthermore, the biofilm disruption and antibiotic potentiation activity identify PgaB as a possible therapeutic agent for treatment of PNAG-dependent biofilm infections.AM-2 University of Toronto, McGill University, Hospital for Sick Children | Presentation | 2018-07-23 | Roland Pfoh, "Dustin J. Little ", François LeMauff, Natalie Bamford, "Christina Notte ", Perrin Baker, "Manita Guragain ", "Howard Robinson ", "Gerald B. Pier ", Mark Nitz, "Rajendar Deora ", Donald Sheppard, Patricia Lynne Howell |
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