A New Sialidase Mechanism: Bacteriophage K1F Endo-sialidase is an Inverting Glycosidase

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2009 May 5

PMID 19411257

Citations 14

Authors

Thomas J Morley

Lisa M Willis

Chris Whitfield

Warren W Wakarchuk

Stephen G Withers

Affiliations

Soon will be listed here.

Abstract

Bacteriophages specific for Escherichia coli K1 express a tailspike protein that degrades the polysialic acid coat of E. coli K1 that is essential for bacteriophage infection. This enzyme is specific for polysialic acid and is a member of a family of endo-sialidases. This family is unusual because all other previously reported sialidases outside of this family are exo- or trans-sialidases. The recently determined structure of an endo-sialidase derived from bacteriophage K1F (endoNF) revealed an active site that lacks a number of the residues that are conserved in other sialidases, implying a new, endo-sialidase-specific catalytic mechanism. Using synthetic trifluoromethylumbelliferyl oligosialoside substrates, kinetic parameters for hydrolysis at a single cleavage site were determined. Measurement of kcat/Km at a series of pH values revealed a dependence on a single protonated group of pKa 5. Mutation of a putative active site acidic residue, E581A, resulted in complete loss of sialidase activity. Direct 1H NMR analysis of the hydrolysis of trifluoromethylumbelliferyl sialotrioside revealed that endoNF is an inverting sialidase. All other wild type sialidases previously reported are retaining glycosidases, implying a new mechanism of sialidase action specific to this family of endo-sialidases.

Citing Articles

Targeting Pseudomonas aeruginosa biofilm with an evolutionary trained bacteriophage cocktail exploiting phage resistance trade-offs.

Kunisch F, Campobasso C, Wagemans J, Yildirim S, Chan B, Schaudinn C Nat Commun. 2024; 15(1):8572.

PMID: 39362854 PMC: 11450229. DOI: 10.1038/s41467-024-52595-w.

Site-specific immobilization of the endosialidase reveals QSOX2 is a novel polysialylated protein.

Hunter C, Derksen T, Makhsous S, Doll M, Perez S, Scott N Glycobiology. 2024; 34(5).

PMID: 38489772 PMC: 11031136. DOI: 10.1093/glycob/cwae026.

Sialidase Inhibitors with Different Mechanisms.

Keil J, Rafn G, Turan I, Aljohani M, Sahebjam-Atabaki R, Sun X J Med Chem. 2022; 65(20):13574-13593.

PMID: 36252951 PMC: 9620260. DOI: 10.1021/acs.jmedchem.2c01258.

-Sialylation: a strategy used to incorporate sialic acid into oligosaccharides.

de Lederkremer R, Giorgi M, Agusti R RSC Chem Biol. 2022; 3(2):121-139.

PMID: 35360885 PMC: 8827155. DOI: 10.1039/d1cb00176k.

Crystal structure of the Propionibacterium acnes surface sialidase, a drug target for P. acnes-associated diseases.

Yu A, Volkers G, Jongkees S, Worrall L, Withers S, Strynadka N Glycobiology. 2021; 32(2):162-170.

PMID: 34792586 PMC: 8934140. DOI: 10.1093/glycob/cwab094.

References

Hildebrandt H, Muhlenhoff M, Weinhold B, Gerardy-Schahn R . Dissecting polysialic acid and NCAM functions in brain development. J Neurochem. 2007; 103 Suppl 1:56-64. DOI: 10.1111/j.1471-4159.2007.04716.x. View

Seidenfaden R, Krauter A, Schertzinger F, Gerardy-Schahn R, Hildebrandt H . Polysialic acid directs tumor cell growth by controlling heterophilic neural cell adhesion molecule interactions. Mol Cell Biol. 2003; 23(16):5908-18. PMC: 166353. DOI: 10.1128/MCB.23.16.5908-5918.2003. View

Watson J, Indurugalla D, Cheng L, Narine A, Bennet A . The hydrolase and transferase activity of an inverting mutant sialidase using non-natural beta-sialoside substrates. Biochemistry. 2006; 45(44):13264-75. DOI: 10.1021/bi061489x. View

Finne J, Leinonen M, Makela P . Antigenic similarities between brain components and bacteria causing meningitis. Implications for vaccine development and pathogenesis. Lancet. 1983; 2(8346):355-7. DOI: 10.1016/s0140-6736(83)90340-9. View

GROSS R, Cheasty T, Rowe B . Isolation of bacteriophages specific for the K1 polysaccharide antigen of Escherichia coli. J Clin Microbiol. 1977; 6(6):548-50. PMC: 274821. DOI: 10.1128/jcm.6.6.548-550.1977. View

Bhattacharjee A, Jennings H, KENNY C, Martin A, Smith I . Structural determination of the sialic acid polysaccharide antigens of Neisseria meningitidis serogroups B and C with carbon 13 nuclear magnetic resonance. J Biol Chem. 1975; 250(5):1926-32. View

Brayer G, Sidhu G, MAURUS R, Rydberg E, Braun C, Wang Y . Subsite mapping of the human pancreatic alpha-amylase active site through structural, kinetic, and mutagenesis techniques. Biochemistry. 2000; 39(16):4778-91. DOI: 10.1021/bi9921182. View

Schauer R . Achievements and challenges of sialic acid research. Glycoconj J. 2001; 17(7-9):485-99. PMC: 7087979. DOI: 10.1023/a:1011062223612. View

Long G, Bryant J, Taylor P, Luzio J . Complete nucleotide sequence of the gene encoding bacteriophage E endosialidase: implications for K1E endosialidase structure and function. Biochem J. 1995; 309 ( Pt 2):543-50. PMC: 1135765. DOI: 10.1042/bj3090543. View

10.

Willis L, Gilbert M, Karwaski M, Blanchard M, Wakarchuk W . Characterization of the alpha-2,8-polysialyltransferase from Neisseria meningitidis with synthetic acceptors, and the development of a self-priming polysialyltransferase fusion enzyme. Glycobiology. 2007; 18(2):177-86. DOI: 10.1093/glycob/cwm126. View

11.

Schauer R . Sialic acids: fascinating sugars in higher animals and man. Zoology (Jena). 2005; 107(1):49-64. DOI: 10.1016/j.zool.2003.10.002. View

12.

Franz C, Rutishauser U, Rafuse V . Polysialylated neural cell adhesion molecule is necessary for selective targeting of regenerating motor neurons. J Neurosci. 2005; 25(8):2081-91. PMC: 6726067. DOI: 10.1523/JNEUROSCI.4880-04.2005. View

13.

Moolenaar C, Muller E, Schol D, Figdor C, Bock E, Bitter-Suermann D . Expression of neural cell adhesion molecule-related sialoglycoprotein in small cell lung cancer and neuroblastoma cell lines H69 and CHP-212. Cancer Res. 1990; 50(4):1102-6. View

14.

Rutishauser U . Polysialic acid at the cell surface: biophysics in service of cell interactions and tissue plasticity. J Cell Biochem. 1998; 70(3):304-12. DOI: 10.1002/(sici)1097-4644(19980901)70:3<304::aid-jcb3>3.0.co;2-r. View

15.

Michon F, Brisson J, Jennings H . Conformational differences between linear alpha (2----8)-linked homosialooligosaccharides and the epitope of the group B meningococcal polysaccharide. Biochemistry. 1987; 26(25):8399-405. DOI: 10.1021/bi00399a055. View

16.

Wang Q, Trimbur D, Graham R, Warren R, Withers S . Identification of the acid/base catalyst in Agrobacterium faecalis beta-glucosidase by kinetic analysis of mutants. Biochemistry. 1995; 34(44):14554-62. DOI: 10.1021/bi00044a034. View

17.

Kao Y, Lerner L, Warner T . Stereoselectivity of the Chinese hamster ovary cell sialidase: sialoside hydrolysis with overall retention of configuration. Glycobiology. 1997; 7(4):559-63. DOI: 10.1093/glycob/7.4.559. View

18.

Finne J, Makela P . Cleavage of the polysialosyl units of brain glycoproteins by a bacteriophage endosialidase. Involvement of a long oligosaccharide segment in molecular interactions of polysialic acid. J Biol Chem. 1985; 260(2):1265-70. View

19.

Manzoni M, Colombi P, Papini N, Rubaga L, Tiso N, Preti A . Molecular cloning and biochemical characterization of sialidases from zebrafish (Danio rerio). Biochem J. 2007; 408(3):395-406. PMC: 2267369. DOI: 10.1042/BJ20070627. View

20.

Petter J, Vimr E . Complete nucleotide sequence of the bacteriophage K1F tail gene encoding endo-N-acylneuraminidase (endo-N) and comparison to an endo-N homolog in bacteriophage PK1E. J Bacteriol. 1993; 175(14):4354-63. PMC: 204875. DOI: 10.1128/jb.175.14.4354-4363.1993. View