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  Oligomerization of the SPP1 scaffolding protein

Poh, S. L., el Khadali, F., Berrier, C., Lurz, R., Melki, R., & Tavares, P. (2008). Oligomerization of the SPP1 scaffolding protein. Journal of Molecular Biology, 378(3), 551-564. doi:scaffolding protein; circular dichroism; chemical cross-linking; procapsid assembly; protein association.

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Item Permalink: https://hdl.handle.net/11858/00-001M-0000-0010-804E-0 Version Permalink: https://hdl.handle.net/11858/00-001M-0000-0010-804F-E
Genre: Journal Article
Alternative Title : J Mol Biol.

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 Creators:
Poh, Siew Lay, Author
el Khadali, Fatima, Author
Berrier, Catherine, Author
Lurz, Rudi1, Author
Melki, Ronald, Author
Tavares, Paulo, Author
Affiliations:
1Max Planck Society, ou_persistent13              

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Free keywords: scaffolding protein; circular dichroism; chemical cross-linking; procapsid assembly; protein association
 Abstract: Viral scaffolding proteins direct polymerization of major capsid protein subunits into icosahedral procapsid structures. The scaffolding protein of bacteriophage SPP1 was engineered with a C-terminal hexahistidine tag (gp11-His6) and purified. The protein is an α-helical-rich molecule with a very elongated shape as found for internal scaffolding proteins from other phages. It is a 3.3 S tetramer of 93.6 kDa at micromolar concentrations. Intersubunit cross-linking of these tetramers generated preferentially covalently bound dimers, revealing that gp11-His6 is structurally a dimer of dimers. Incubation at temperatures above 37 °C correlated with a reduction of its α-helical content and a less effective intersubunit cross-linking. Complete loss of secondary structure was observed at temperatures above 60 °C. Refolding of gp11-His6 thermally denatured at 65 °C led to reacquisition of the protein native ellipticity spectrum but the resulting population of molecules was heterogeneous. Its hydrodynamic behavior was compatible with a mix of 3.3 S elongated tetramers (not, vert, similar 90%) and a smaller fraction of 2.4 S dimers (not, vert, similar 10%). This population of gp11-His6 was competent to direct polymerization of the SPP1 major capsid protein gp13 into procapsid-like structures in a newly developed assembly assay in vitro. Although native tetramers were active in assembly, refolded gp11-His6 showed enhanced binding to gp13 revealing a more active species for interaction with the major capsid protein than native gp11-His6.

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Language(s): eng - English
 Dates: 2008-02-23
 Publication Status: Issued
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: eDoc: 412083
URI: http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WK7-4RX079N-1&_user=28761&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000002718&_version=1&_urlVersion=0&_userid=28761&md5=32d1f30d71dacd605d42120622dbba53
DOI: scaffolding protein; circular dichroism; chemical cross-linking; procapsid assembly; protein association
 Degree: -

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Title: Journal of Molecular Biology
  Alternative Title : J Mol Biol.
Source Genre: Journal
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Pages: - Volume / Issue: 378 (3) Sequence Number: - Start / End Page: 551 - 564 Identifier: -