Streptococcal phosphotransferase system imports unsaturated hyaluronan disaccharide derived from host extracellular matrices

Certain bacterial species target the polysaccharide glycosaminoglycans (GAGs) of animal extracellular matrices for colonization and/or infection. GAGs such as hyaluronan and chondroitin sulfate consist of repeating disaccharide units of uronate and amino sugar residues, and are depolymerized to unsaturated disaccharides by bacterial extracellular or cell-surface polysaccharide lyase. The disaccharides are degraded and metabolized by cytoplasmic enzymes such as unsaturated glucuronyl hydrolase, isomerase, and reductase. The genes encoding these enzymes are assembled to form a GAG genetic cluster. Here, we demonstrate the Streptococcus agalactiae phosphotransferase system (PTS) for import of unsaturated hyaluronan disaccharide. S. agalactiae NEM316 was found to depolymerize and assimilate hyaluronan, whereas its mutant with a disruption in PTS genes included in the GAG cluster was unable to grow on hyaluronan, while retaining the ability to depolymerize hyaluronan. Using toluene-treated wild-type cells, the PTS import activity of unsaturated hyaluronan disaccharide was significantly higher than that observed in the absence of the substrate. In contrast, the PTS mutant was unable to import unsaturated hyaluronan disaccharide, indicating that the corresponding PTS is the only importer of fragmented hyaluronan, which is suitable for PTS to phosphorylate the substrate at the C-6 position. The three-dimensional structure of streptococcal EIIA, one of the PTS components, was found to contain a Rossman-fold motif by X-ray crystallization. Docking of EIIA with another component EIIB by modeling provided structural insights into the phosphate transfer mechanism. This study is the first to identify the substrate (unsaturated hyaluronan disaccharide) recognized and imported by the streptococcal PTS. IMPORTANCE (118/120 words) The PTS identified in this work imports sulfate group-free unsaturated hyaluronan disaccharide as a result of the phosphorylation of the substrate at the C-6 position. S. agalactiae can be indigenous to animal hyaluronan-rich tissues owing to the bacterial molecular system for fragmentation, import, degradation, and metabolism of hyaluronan. Distinct from hyaluronan, most GAGs, which are sulfated at the C-6 position, are unsuitable for PTS due to its inability to phosphorylate the substrate. More recently, we have identified a solute-binding protein-dependent ABC transporter in a pathogenic Streptobacillus moniliformis as an importer of sulfated and non-sulfated fragmented GAGs without any substrate modification. Our findings regarding PTS and ABC transporter shed light on bacterial clever colonization/infection system targeting various animal GAGs.

[1]  R. Markwald,et al.  Chapter 1 Introduction to COVID-19 , 2022, COVID-19 in the Environment.

[2]  S. Ricard-Blum,et al.  Proteoglycan Chemical Diversity Drives Multifunctional Cell Regulation and Therapeutics. , 2018, Chemical reviews.

[3]  K. Murata,et al.  Probiotics in human gut microbiota can degrade host glycosaminoglycans , 2018, Scientific Reports.

[4]  Torsten Schwede,et al.  SWISS-MODEL: homology modelling of protein structures and complexes , 2018, Nucleic Acids Res..

[5]  B. Mikami,et al.  Alternative substrate-bound conformation of bacterial solute-binding protein involved in the import of mammalian host glycosaminoglycans , 2017, Scientific Reports.

[6]  B. Mikami,et al.  A bacterial ABC transporter enables import of mammalian host glycosaminoglycans , 2017, Scientific Reports.

[7]  Mikko Arvas,et al.  A novel pathway for fungal D-glucuronate catabolism contains an L-idonate forming 2-keto-L-gulonate reductase , 2016, Scientific Reports.

[8]  B. Mikami,et al.  Metabolic Fate of Unsaturated Glucuronic/Iduronic Acids from Glycosaminoglycans , 2015, The Journal of Biological Chemistry.

[9]  D. Zurawski,et al.  Staphylococcus aureus Hyaluronidase Is a CodY-Regulated Virulence Factor , 2014, Infection and Immunity.

[10]  R. Linhardt,et al.  Masquerading microbial pathogens: capsular polysaccharides mimic host-tissue molecules. , 2014, FEMS microbiology reviews.

[11]  R. Burne,et al.  Uptake and Metabolism of N-Acetylglucosamine and Glucosamine by Streptococcus mutans , 2014, Applied and Environmental Microbiology.

[12]  J. Deutscher,et al.  The Bacterial Phosphoenolpyruvate:Carbohydrate Phosphotransferase System: Regulation by Protein Phosphorylation and Phosphorylation-Dependent Protein-Protein Interactions , 2014, Microbiology and Molecular Reviews.

[13]  Chengping Lu,et al.  Two Novel Functions of Hyaluronidase from Streptococcus agalactiae Are Enhanced Intracellular Survival and Inhibition of Proinflammatory Cytokine Expression , 2014, Infection and Immunity.

[14]  D. Leppert,et al.  Matrix Metalloproteinase Inhibition Lowers Mortality and Brain Injury in Experimental Pneumococcal Meningitis , 2014, Infection and Immunity.

[15]  R. Linhardt,et al.  Glycosaminoglycans in infectious disease , 2013, Biological reviews of the Cambridge Philosophical Society.

[16]  S. King,et al.  Streptococcus pneumoniae Can Utilize Multiple Sources of Hyaluronic Acid for Growth , 2012, Infection and Immunity.

[17]  H. Abriouel,et al.  Enterococci as probiotics and their implications in food safety. , 2011, International journal of food microbiology.

[18]  O. Kuipers,et al.  CelR-mediated activation of the cellobiose-utilization gene cluster in Streptococcus pneumoniae. , 2011, Microbiology.

[19]  Valerie M. Weaver,et al.  The extracellular matrix at a glance , 2010, Journal of Cell Science.

[20]  B. Mikami,et al.  Structural Determinants in Streptococcal Unsaturated Glucuronyl Hydrolase for Recognition of Glycosaminoglycan Sulfate Groups* , 2010, The Journal of Biological Chemistry.

[21]  Liisa Holm,et al.  Dali server: conservation mapping in 3D , 2010, Nucleic Acids Res..

[22]  P. Emsley,et al.  Features and development of Coot , 2010, Acta crystallographica. Section D, Biological crystallography.

[23]  P. Nyirjesy,et al.  Hyaluronan in vaginal secretions: association with recurrent vulvovaginal candidiasis. , 2009, American journal of obstetrics and gynecology.

[24]  B. Mikami,et al.  Substrate Specificity of Streptococcal Unsaturated Glucuronyl Hydrolases for Sulfated Glycosaminoglycan* , 2009, The Journal of Biological Chemistry.

[25]  Neha S. Gandhi,et al.  The Structure of Glycosaminoglycans and their Interactions with Proteins , 2008, Chemical biology & drug design.

[26]  G. Clore,et al.  Solution NMR Structures of Productive and Non-productive Complexes between the A and B Domains of the Cytoplasmic Subunit of the Mannose Transporter of the Escherichia coli Phosphotransferase System* , 2008, Journal of Biological Chemistry.

[27]  K. Henrick,et al.  Inference of macromolecular assemblies from crystalline state. , 2007, Journal of molecular biology.

[28]  M. J. Jedrzejas Unveiling molecular mechanisms of bacterial surface proteins: Streptococcus pneumoniae as a model organism for structural studies , 2007, Cellular and Molecular Life Sciences.

[29]  U. Schumacher,et al.  Hyaluronan Export by the ABC Transporter MRP5 and Its Modulation by Intracellular cGMP* , 2007, Journal of Biological Chemistry.

[30]  B. Mikami,et al.  Crystal Structure of Unsaturated Glucuronyl Hydrolase Complexed with Substrate , 2006, Journal of Biological Chemistry.

[31]  M. J. Jedrzejas,et al.  Hyaluronidases: their genomics, structures, and mechanisms of action. , 2006, Chemical reviews.

[32]  M. Saier,et al.  Comparative Genomic Analyses of the Bacterial Phosphotransferase System , 2005, Microbiology and Molecular Biology Reviews.

[33]  M. Mulholland,et al.  Classification of chondroitin sulfate A, chondroitin sulfate C, glucosamine hydrochloride and glucosamine 6 sulfate using chemometric techniques. , 2005, Journal of pharmaceutical and biomedical analysis.

[34]  G Marius Clore,et al.  Solution NMR Structure of the 48-kDa IIAMannose-HPr Complex of the Escherichia coli Mannose Phosphotransferase System* , 2005, Journal of Biological Chemistry.

[35]  M. Saier,et al.  Evolution of the bacterial phosphotransferase system: from carriers and enzymes to group translocators. , 2005, Biochemical Society transactions.

[36]  B. Spellerberg,et al.  Hyaluronan release from Streptococcus pyogenes: export by an ABC transporter. , 2004, Glycobiology.

[37]  J. Goodwin,et al.  University of Texas Medical Branch at Galveston , 2004, Academic medicine : journal of the Association of American Medical Colleges.

[38]  Jue Chen,et al.  ATP-binding cassette transporters in bacteria. , 2004, Annual review of biochemistry.

[39]  Carmen Buchrieser,et al.  Genome sequence of Streptococcus agalactiae, a pathogen causing invasive neonatal disease , 2002, Molecular microbiology.

[40]  Songlin Li,et al.  Hyaluronan Binding and Degradation byStreptococcus agalactiae Hyaluronate Lyase* , 2001, The Journal of Biological Chemistry.

[41]  M. J. Jedrzejas Pneumococcal Virulence Factors: Structure and Function , 2001, Microbiology and Molecular Biology Reviews.

[42]  O Habuchi,et al.  Diversity and functions of glycosaminoglycan sulfotransferases. , 2000, Biochimica et biophysica acta.

[43]  K. Prydz,et al.  Synthesis and sorting of proteoglycans. , 2000, Journal of cell science.

[44]  S. Kawai,et al.  Unsaturated glucuronyl hydrolase of Bacillus sp. GL1: novel enzyme prerequisite for metabolism of unsaturated oligosaccharides produced by polysaccharide lyases. , 1999, Archives of biochemistry and biophysics.

[45]  A. Vagin,et al.  MOLREP: an Automated Program for Molecular Replacement , 1997 .

[46]  H. Yu,et al.  Specificity of the hyaluronate lyase of group-B streptococcus toward unsulphated regions of chondroitin sulphate. , 1997, The Biochemical journal.

[47]  B. Erni,et al.  Structure of the IIA domain of the mannose transporter from Escherichia coli at 1.7 angstroms resolution. , 1996, Journal of molecular biology.

[48]  D. Sawitzky Protein-glycosaminoglycan interactions: infectiological aspects , 1996, Medical Microbiology and Immunology.

[49]  S. Grzesiek,et al.  NMRPipe: A multidimensional spectral processing system based on UNIX pipes , 1995, Journal of biomolecular NMR.

[50]  M. L. Ricci,et al.  Electrotransformation of Streptococcus agalactiae with plasmid DNA. , 1994, FEMS microbiology letters.

[51]  J. Scott Supramolecular organization of extracellular matrix glycosaminoglycans, in vitro and in the tissues , 1992, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[52]  T. Laurent,et al.  Localization of hyaluronan in regions of the human female reproductive tract. , 1991, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[53]  P. K. Smith,et al.  Measurement of protein using bicinchoninic acid. , 1985, Analytical biochemistry.

[54]  P. Postma,et al.  Phosphoenolpyruvate:carbohydrate phosphotransferase system of bacteria , 1985 .

[55]  P. Postma,et al.  Phosphoenolpyruvate:carbohydrate phosphotransferase system of bacteria. , 1985, Microbiological reviews.

[56]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[57]  M. Saier Bacterial phosphoenolpyruvate: sugar phosphotransferase systems: structural, functional, and evolutionary interrelationships. , 1977, Bacteriological reviews.

[58]  G. Shockman,et al.  Growth of several cariogenic strains of oral streptococci in a chemically defined medium , 1975, Infection and immunity.

[59]  H. Kornberg,et al.  Inducible phosphoenolpyruvate-dependent hexose phosphotransferase activities in Escherichia coli. , 1972, The Biochemical journal.

[60]  S. Roseman,et al.  Genetic evidence for the role of a bacterial phosphotransferase system in sugar transport. , 1967, Proceedings of the National Academy of Sciences of the United States of America.

[61]  M. Mathews,et al.  The determination of chondroitin sulfate C-type polysaccharides in mixtures with other acid mucopolysaccharides. , 1961, Biochimica et biophysica acta.

[62]  W. Delano The PyMOL Molecular Graphics System , 2002 .

[63]  Vincent B. Chen,et al.  Acta Crystallographica Section D Biological , 2001 .

[64]  Z. Otwinowski,et al.  Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[65]  Z. Otwinowski,et al.  [20] Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[66]  R. Langer,et al.  Enzymatic degradation of glycosaminoglycans. , 1995, Critical reviews in biochemistry and molecular biology.