Bacteriophage P22 scaffolding protein forms oligomers in solution

Journal of Molecular Biology
M H ParkerP E Prevelige

Abstract

The scaffolding protein of Salmonella typhimurium bacteriophage P22 is a 33.6 kDa protein required both in vivo and in vitro for the polymerization of the viral coat protein into closed T = 7 icosahedral procapsids. In vitro assembly reaction kinetics have previously been found to vary between second and third order with respect to scaffolding protein concentration, suggesting that dimers and/or higher-order oligomers may be the active species in assembly. Analytical ultracentrifugation experiments suggest that scaffolding protein undergoes a rapidly-reversible monomer/dimer/tetramer equilibrium, with higher association constants at 4 degrees C than at 20 degrees C. Under conditions in which in vitro assembly reactions are carried out (30 to 1000 microg/ml scaffolding protein, 20 degrees C), monomers are the predominant species, but the concentration of dimers is significant. A mutant scaffolding protein, R74C/L177I, which forms disulfide-linked dimers, catalyzed procapsid assembly at a higher rate than did the wild-type scaffolding protein; preincubation in dithiothreitol had little effect on the wild-type protein, but greatly reduced the activity of the mutant. These findings suggest that dimers and/or higher-order oligomers ...Continue Reading

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Citations

Jan 12, 2011·Proceedings of the National Academy of Sciences of the United States of America·Dong-Hua ChenWah Chiu
Aug 11, 2012·The Journal of Biological Chemistry·G Pauline Padilla-MeierSherwood R Casjens
Jul 14, 2017·Nanoscale·Jhanvi SharmaTrevor Douglas
Mar 3, 2018·Chemical Society Reviews·William M AumillerTrevor Douglas
May 29, 1999·The Journal of Biological Chemistry·B Greene, J King
Jan 24, 2008·Chembiochem : a European Journal of Chemical Biology·Sebyung KangPeter E Prevelige
Jul 29, 1998·Journal of Molecular Biology·R TumaP E Prevelige
Jul 29, 1998·Journal of Molecular Biology·M H ParkerP E Prevelige
Feb 6, 2002·Journal of Molecular Biology·Sean D Moore, Peter E Prevelige
Sep 16, 2003·Journal of Molecular Biology·Shannon M DoyleCarolyn M Teschke
Apr 22, 2005·Journal of Molecular Biology·Peter R WeigeleSherwood R Casjens
Aug 11, 2018·Biomacromolecules·Kimberly McCoyTrevor Douglas
Apr 2, 2008·Journal of Molecular Biology·Siew Lay PohPaulo Tavares
May 25, 2011·Journal of Molecular Biology·G Pauline Padilla-Meier, Carolyn M Teschke

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