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A genetic analysis of an important hydrophobic interaction at the P22 tailspike protein N-terminal domain

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Abstract

P22 bacteriophage has been studied extensively and has served as a model for many important processes such as in vivo protein folding, protein aggregation and protein-protein interactions. The trimeric tailspike protein (TSP) serves as the receptor-binding protein for the P22 bacteriophage to the bacterial host. The homotrimeric P22 tail consists of three chains of 666aa in which the first 108aa form a trimeric dome-like structure which is called the N-terminal domain (NTD) and is responsible for attachment of the tailspike protein to the rest of the phage particle structure in the phage assembly pathway. Knowledge of this interaction requires information on what amino acids are interacting in the interface and how the NTD structure is maintained. The first 23aa form the “stem peptide” which originates at the dome top and terminates at the dome bottom. It contains a hydrophobic valine patch (V8-V9-V10) located within the dome structure. It is hypothesized that the interaction between the hydrophobic valine patch located on stem peptide and the adjacent polypeptide is critical for the interchain interaction which should be important for the stability of the P22 TSP NTD itself. To test this hypothesis, each amino acid in the valine residues is substituted by an acid, a basic, and a hydrophobic amino acid. The results of such substitutions are presented as well as associated studies. The data strongly suggest that the valine patch is of critical importance in the hydrophobic interaction between stem peptide valine patch and an adjacent chain.

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References

  1. Nobrega FL, Costa AR, Kluskens LD, Azeredo J (2015) Revisiting phage therapy: new applications for old resources. Trends Microbiol 23:185–191

    Article  PubMed  CAS  Google Scholar 

  2. Prevelige Jr PE (2006) Bacteriophage P22. In: Calendar R (ed) The bacteriophages. Oxford Press, New York, pp. 457–468

  3. Switt AIM, Sulakvelidze A, Weidmann M, Kropinski AM, Wishart DS, Poppe C, Liang Y (2015) Salmonella phages and prophages: genomics, taxonomy, and applied aspects. Methods Mol Biol 1225:237–287

    Article  PubMed  CAS  Google Scholar 

  4. Aksyuk AA, Rossmann MG (2011) Bacteriophage assembly. Viruses 3:172–203

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  5. Domitrovic T, Movahed N, Bothner B, Matsui T, Wang Q, Doerschuk P, Johnson J (2013) Virus assembly and maturation: auto-regulation through allosteric molecular switches. J Mol Biol 425:1488–1496

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. Sauer RT, Krovatin W, Poteete AR, Berget PB (1982) Phage P22 tail protein: gene and amino acid sequence. Biochemistry 21:5811–5815

    Article  PubMed  CAS  Google Scholar 

  7. Seul A, Muller JJ, Andres D, Stettner E, Heinemann U, Seckler R (2014) Bacteriophage P22 tailspike: structure of the complete protein and the function of interdomain linker. Acta Cryst 70:1336–1345

    CAS  Google Scholar 

  8. Weigele PR, Scanlon E, King J (2003) Homotrimeric, ß-Stranded viral adhesins and tail proteins. J Bacteriol 185:4022–4030

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  9. Goldenberg D, Berget P, King J (1982) Maturation of the tail spike endorhamnosidase of Salmonella Phage P22. J Biol Chem 257:7864–7871

    PubMed  CAS  Google Scholar 

  10. Poteete AR (1988) The Bacteriophage P22. In: Calendar R (ed) The Bacteriophages. Plenum Press, New York, pp 647–682

    Chapter  Google Scholar 

  11. Steinbacher S, Seckler R, Miller S, Steipe B, Huber R, Reinemer P (1994) Crystal structure of P22 tailspike protein: interdigitated subunits in a thermostable trimer. Science 265:383–386

    Article  PubMed  CAS  Google Scholar 

  12. Steinbacher S, Miller S, Baxa U, Budisa N, Weintraub A, Seckler R, Huber R (1997) Phage P22 tailspike protein: crystal structure of the head-binding domain at 2.3 Ǻ, fully refined structure of the endorhamnosidase at 1.56 Ǻ resolution, and the molecular basis of O-antigen recognition and cleavage. J Mol Biol 267:865–880

    Article  PubMed  CAS  Google Scholar 

  13. Baxa U, Steinbacher S, Miller S, Weintraub A, Huber R, Seckler R (1996) Interactions of phage P22 tails with their cellular receptors. Salmonella o-Antigen Polysacch Biophys J 71:2040–2048

    CAS  Google Scholar 

  14. Palmer C, Williams J, Dean D, Johnson S, Wu H, Jackson D, Villafane R (2014) Stem mutants in the N-terminal domain of the phage P22 tailspike protein. Am J Microbiol Res 2:1–7

    Article  CAS  Google Scholar 

  15. Keskin O, Gursoy A, Ma B, Nussinov R (2008) Principles of protein–protein interactions: what are the preferred ways for proteins to interact? Chem Rev 108:1225–1244

    Article  PubMed  CAS  Google Scholar 

  16. Villafane R, Costa S, Ahmed R, Salgado C (2005) Conservation of the N-terminus of some phage tail proteins. Arch Virol 150:2609–2621

    Article  PubMed  CAS  Google Scholar 

  17. Salgado CJ, Zayas M, Villafane R (2004) Homology between two different Salmonella phages: Salmonella enterica serovar Typhimurium phage P22 and Salmonella enterica serovar Anatum var. 15+phage ε34. Virus Genes 29:87–98

    Article  PubMed  CAS  Google Scholar 

  18. Mateu MG (2009) The capsid protein of human immunodeficiency virus: intersubunit interactions during virus assembly. FEBS J 276:6098–6109

    Article  PubMed  CAS  Google Scholar 

  19. Finzi D, Dieffenbach CW, Basavappa R (2007) Defining and solving the essential protein–protein interactions in HIV infections. J Struct Biol 158:148–155

    Article  PubMed  CAS  Google Scholar 

  20. Wang W-H, Chang L-K, Liu S-T (2011) Molecular interactions of Epstein-Barr capsid proteins. J Virol 85:1618–1624

    Google Scholar 

  21. Robinson A, King J (1997) Disulphide-bonded intermediate on the folding and assembly pathway of a non-disulphide bonded protein. Nat Struct Mol Biol 4:450–455

    Article  CAS  Google Scholar 

  22. Kamei DT, Liu C, Haase-Pettingell C, King JA, Wang DIC, Blandkschtein D (2002) Understanding viral partitioning in two-phase aqueous nonionic micellar systems: 2. Role of attractive interactions between viruses and micelles. Biotech Bioeng 78:190–202

    Article  CAS  Google Scholar 

  23. Schwarz J, Berget P (1989) The isolation and sequence of missense and nonsense mutations in the cloned bacteriophage P22 tailspike protein gene. Genetics 121:635–649

    PubMed  PubMed Central  CAS  Google Scholar 

  24. Betts S, Speed M, King J (1999) Detection of early aggregation intermediates by native gel electrophoresis and native western blotting. Methods Enzymol 309:333–350

    Article  PubMed  CAS  Google Scholar 

  25. Roskams J, Rodgers L (2002) Electrophoretic separation of proteins and nucleic acids. In: Roskams J, Rodgers L (eds) Lab Ref: A handbook of recipes, reagents, and other reference tools for use at the bench. Cold Spring Harbor Press, New York, pp 63–91

    Google Scholar 

  26. Maurides PA, Schwarz JJ, Berget PB (1990) Intragenic suppression of a capsid assembly-defective P22 tailspike mutation. Genetics 125:673–681

    PubMed  PubMed Central  CAS  Google Scholar 

  27. Chen B, King J (1991) Thermal unfolding pathway for the thermostable P22 tailspike endorhamnosidase. Biochemistry 30:6260–6269

    Article  PubMed  CAS  Google Scholar 

  28. Sturtevant J, Yu M, Haase-Pettingell C, King J (1989) Thermostability of temperature-sensitive folding mutants of the P22 tailspike protein. J Biol Chem 264:10693–10698

    PubMed  CAS  Google Scholar 

  29. Burley S, Petsko G (1985) Aromatic-aromatic interaction: a mechanism of protein structure stabilization. Science 229:23–28. https://doi.org/10.1126/science.3892686

    Article  PubMed  CAS  Google Scholar 

  30. Kessel A, Ben-Tal N (2011) Introduction to proteins, 1st edn. CRC Press, Boca Raton, pp 44–58

    Google Scholar 

  31. Keskin O, Gursoy A, Ma B, Nussinov R (2008) Principles of protein–protein interactions: what are the preferred ways for proteins to interact? Chem Rev 108:1225–1244

    Article  PubMed  CAS  Google Scholar 

  32. Prevost M, Wodak S, Tidor B, Karplus M (1991) Contribution of the hydrophobic effect to protein stability: analysis based on simulations of the Ile-96—Ala mutation in barnase. PNAS 88:10880–10884

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  33. Manning M, Colón W (2004) Structural basis of protein kinetic stability: resistance to sodium dodecyl sulfate suggests a central role for rigidity and a bias toward β-sheet structure. Biochemistry 43:11248–11254

    Article  PubMed  CAS  Google Scholar 

  34. Xia K, Zhang S, Bathrick B, Liu S, Garcia Y, Colón W (2012) Quantifying the kinetic stability of hyperstable proteins via time-dependent SDS trapping. Biochemistry 51:100–107

    Article  PubMed  CAS  Google Scholar 

  35. Freiberg A, Morona R, Van Den Bosch L, Jung C, Behlke J, Carlin N, Seckler R, Baxa U (2002) The Tailspike Protein of STIgella Phage Sf6: a structural homolog of salmonella phage p22 tailspike protein without sequence similarity in the β-helix domain. J Biol Chem 278(3):1542–1548

    Article  PubMed  CAS  Google Scholar 

  36. Wen J, Arthur K, Chemmalil L, Muzammil S, Gabrielson J, Jiang Y (2012) Applications of can differential scanning calorimetry for thermal stability analysis of proteins: qualification of DSC. J Pharmaceut Sci 101:955–964

    Article  CAS  Google Scholar 

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Acknowledgements

These studies were supported by the Deanships of the College of Science, Mathematics and Technology at the Alabama State University and by the Department of Chemistry and Biochemistry at Huntingdon College. The authors also thank the anonymous reviewers for insightful comments.

Funding

This work was funded in part by Deanship of the College of Science, Technology, Engineering and Mathematics (C-STEM) from Alabama State University.

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Authors and Affiliations

  1. Program in Microbiology, Alabama State University, 1627 Harris Way, Montgomery, AL, 36104, USA

    Jeremie Williams, Karthikeya Venkatesan, Joseph Atia Ayariga, Hongzhuan Wu & Robert Villafane

  2. Department of Chemistry and Biochemistry, Huntingdon College, Montgomery, AL, USA

    Doba Jackson

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  1. Jeremie Williams
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  2. Karthikeya Venkatesan
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  3. Joseph Atia Ayariga
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  4. Doba Jackson
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  5. Hongzhuan Wu
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  6. Robert Villafane
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Correspondence to Robert Villafane.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Handling Editor: T. K. Frey.

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Williams, J., Venkatesan, K., Ayariga, J.A. et al. A genetic analysis of an important hydrophobic interaction at the P22 tailspike protein N-terminal domain. Arch Virol 163, 1623–1633 (2018). https://doi.org/10.1007/s00705-018-3777-y

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  • DOI: https://doi.org/10.1007/s00705-018-3777-y

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