C-terminal hydrophobic interactions play a critical role in oligomeric assembly of the P22 tailspike trimer

Matthew J Gage, Anne Skaja Robinson

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The tailspike protein from the bacteriophage P22 is a well characterized model system for folding and assembly of multimeric proteins. Folding intermediates from both the in vivo and in vitro pathways have been identified, and both the initial folding steps and the protrimer-to-trimer transition have been well studied. In contrast, there has been little experimental evidence to describe the assembly of the protrimer. Previous results indicated that the C terminus plays a critical role in the overall stability of the P22 tailspike protein. Here, we present evidence that the C terminus is also the critical assembly point for trimer assembly. Three truncations of the full-length tailspike protein, TSPΔN, TSPΔC, and TSPΔNC, were generated and tested for their ability to form mixed trimer species. TSPΔN forms mixed trimers with full-length P22 tailspike, but TSPΔC and TSPΔNC are incapable of forming similar mixed trimer species. In addition, mutations in the hydrophobic core of the C terminus were unable to form trimer in vivo. Finally, the hydrophobic-binding dye ANS inhibits the formation of trimer by inhibiting progression through the folding pathway. Taken together, these results suggest that hydrophobic interactions between C-terminal regions of P22 tailspike monomers play a critical role in the assembly of the P22 tailspike trimer.

Original languageEnglish (US)
Pages (from-to)2732-2747
Number of pages16
JournalProtein Science
Volume12
Issue number12
DOIs
StatePublished - Dec 2003
Externally publishedYes

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Hydrophobic and Hydrophilic Interactions
Proteins
Bacteriophage P22
Coloring Agents
Mutation
Monomers

Keywords

  • ANS
  • Assembly
  • Folding
  • Hydrophobic interactions
  • P22 tailspike

ASJC Scopus subject areas

  • Biochemistry

Cite this

C-terminal hydrophobic interactions play a critical role in oligomeric assembly of the P22 tailspike trimer. / Gage, Matthew J; Robinson, Anne Skaja.

In: Protein Science, Vol. 12, No. 12, 12.2003, p. 2732-2747.

Research output: Contribution to journalArticle

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