The tailspike protein of Shigella phage Sf6: A structural homolog of Salmonella phage P22 tailspike protein without sequence similarity in the β-helix domain

Alexander Freiberg, Renato Morona, Luisa Van den Bosch, Christiane Jung, Joachim Behlke, Nils Carlin, Robert Seckler, Ulrich Baxa

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36 Scopus citations

Abstract

Bacteriophage Sf6 tailspike protein is functionally equivalent to the well characterized tailspike of Salmonella phage P22, mediating attachment of the viral particle to host cell-surface polysaccharide. However, there is significant sequence similarity between the two 70-kDa polypeptides only in the N-terminal putative capsid-binding domains. The major, central part of P22 tail-spike protein, which forms a parallel β-helix and is responsible for saccharide binding and hydrolysis, lacks detectable sequence homology to the Sf6 protein. After recombinant expression in Escherichia coli as a soluble protein, the Sf6 protein was purified to homogeneity. As shown by circular dichroism and Fourier transform infrared spectroscopy, the secondary structure contents of Sf6 and P22 tailspike proteins are very similar. Both tailspikes are thermostable homotrimers and resist denaturation by SDS at room temperature. The specific endorhamnosidase activities of Sf6 tailspike protein toward fluorescence-labeled dodeca-, deca-, and octasaccharide fragments of Shigella O-antigen suggest a similar active site topology of both proteins. Upon deletion of the N-terminal putative capsid-binding domain, the protein still forms a thermostable, SDS-resistant trimer that has been crystallized. The observations strongly suggest that the tailspike of phage Sf6 is a trimeric parallel β-helix protein with high structural similarity to its functional homolog from phage P22.

Original languageEnglish (US)
Pages (from-to)1542-1548
Number of pages7
JournalJournal of Biological Chemistry
Volume278
Issue number3
DOIs
StatePublished - Jan 17 2003
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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