PMID: 12767837May 28, 2003Paper

The folding mechanism of a two-domain protein: folding kinetics and domain docking of the gene-3 protein of phage fd

Journal of Molecular Biology
Andreas Martin, Franz X Schmid

Abstract

The gene-3 protein (G3P) of filamentous phages is essential for the infection of Escherichia coli. The carboxy-terminal domain anchors this protein in the phage coat, whereas the two amino-terminal domains N1 and N2 protrude from the phage surface. We analyzed the folding mechanism of the two-domain fragment N1-N2 of G3P (G3P(*)) and the interplay between folding and domain assembly. For this analysis, a variant of G3P(*) was used that contained four stabilizing mutations (IIHY-G3P(*)). The observed refolding kinetics extend from 10 ms to several hours. Domain N1 refolds very rapidly (with a time constant of 9.4 ms at 0.5 M guanidinium chloride, 25 degrees C) both as a part of IIHY-G3P(*) and as an isolated protein fragment. The refolding of domain N2 is slower and involves two reactions with time constants of seven seconds and 42 seconds. These folding reactions of the individual domains are followed by a very slow, spectroscopically silent docking process, which shows a time constant of 6200 seconds. This reaction was detected by a kinetic unfolding assay for native molecules. Before docking, N1 and N2 unfold fast and independently, after docking they unfold slowly in a correlated fashion. A high energy barrier is thus create...Continue Reading

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Citations

Dec 31, 2003·Journal of Molecular Biology·Traci B ToppingLisa M Gloss
Aug 21, 2003·Journal of Molecular Biology·Andreas Martin, Franz X Schmid
Jun 7, 2005·Nature Structural & Molecular Biology·Barbara EckertFranz X Schmid
Jul 21, 2009·Proceedings of the National Academy of Sciences of the United States of America·Ulrich WeiningerJochen Balbach
Nov 16, 2010·Protein Engineering, Design & Selection : PEDS·Hauke LilieThomas Kriegel
Mar 10, 2007·Microbiology and Molecular Biology Reviews : MMBR·Eric CascalesDanièle Cavard
Nov 17, 2006·Proceedings of the National Academy of Sciences of the United States of America·Sarah Batey, Jane Clarke
Mar 15, 2013·The Journal of Biological Chemistry·Stephanie Hoffmann-ThomsFranz X Schmid
Mar 8, 2008·Journal of Molecular Biology·Roman P Jakob, Franz X Schmid
Jan 3, 2006·Biophysical Journal·Sarah BateyJane Clarke
Dec 30, 2014·Biochimica Et Biophysica Acta·Philipp A M SchmidpeterFranz X Schmid
Nov 30, 2010·Journal of Molecular Biology·Stefan H LorenzFranz X Schmid
Jan 21, 2014·Journal of Molecular Biology·Stephanie Hoffmann-ThomsFranz X Schmid
Feb 14, 2015·Journal of Molecular Biology·Philipp A M Schmidpeter, Franz X Schmid
Dec 24, 2005·Journal of Molecular Biology·Nicholas J Bennett, Jasna Rakonjac
Mar 12, 2011·Molecular Microbiology·Stefan H Lorenz, Franz X Schmid
May 18, 2012·Molecular Microbiology·Roman P JakobFranz X Schmid
Nov 10, 2005·Chemical Reviews·John W Kehoe, Brian K Kay
Jan 13, 2009·Journal of Molecular Biology·Gabriel ZoldákFranz X Schmid

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