Stability and folding of the cell cycle regulatory protein, p13 suc1 1 1Edited by F. E. Cohen

Rousseau, Frédéric; Schymkowitz, Joost; Pino, Manuel M. Sánchez del; Itzhaki, Laura S.

doi:10.1006/jmbi.1998.2173

Cited by 33 publications

(36 citation statements)

References 72 publications

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“…Mutants were constructed and proteins expressed and purified as described previously (15,16). The samples were pure as judged by SDS͞PAGE and mass spectrometry.…”

Section: Methodsmentioning

confidence: 99%

Three-dimensional domain swapping in p13suc1 occurs in the unfolded state and is controlled by conserved proline residues

Rousseau¹,

Schymkowitz

Wilkinson

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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CorrectionsBIOCHEMISTRY. For the article ''Differential effects of a centrally acting fatty acid synthase inhibitor in lean and obese mice,'' by Monica V. Kumar, Teruhiko Shimokawa, Tim R. Nagy, and M. Daniel Lane, which appeared in number 4, February 19, 2002, of Proc. Natl. Acad. Sci. USA (99, 1921-1925, the authors note the following. ''Under a licensing agreement between FASgen, Inc., and The Johns Hopkins University, Dr. Lane is entitled to a share of royalty received by the University on sales of products that embody the technology described in this article. (98, 10687-10691; First Published August 28, 2001; 10.1073͞pnas.181354398), the authors note the following. In the Introduction and Discussion of our paper, we failed to reference a recent article by Rousseau et al.(1), which demonstrated that single point mutations can significantly perturb the equilibrium between monomeric and domain-swapped dimeric p13suc1. Rational methods were used to redesign p13suc1 from a fully monomeric protein (dissociation constant of Ϸ900 mM) to a fully dimeric protein (dissociation constant of Ϸ100 nM). Fig. 3 appeared incorrectly. The correct version of the figure and its legend appear below.

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“…Mutants were constructed and proteins expressed and purified as described previously (15,16). The samples were pure as judged by SDS͞PAGE and mass spectrometry.…”

Section: Methodsmentioning

confidence: 99%

Three-dimensional domain swapping in p13suc1 occurs in the unfolded state and is controlled by conserved proline residues

Rousseau¹,

Schymkowitz

Wilkinson

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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193

216

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“…5A) was proposed previously to arise from a second monomeric intermediate that must expand to get back to the major pathway to reach the monomeric native state (13). However, there is a marked protein concentration dependence of the refolding reaction above 5 M, suggesting that oligomerization occurs during refolding of suc1 (Fig.…”

Section: An Integrated Model For Folding and Domain Swapping In Suc1-mentioning

confidence: 80%

“…Protein Expression and Purification-The proteins were expressed in Escherichia coli and purified as described previously (13)(14)(15)(16). Briefly, the harvested cells were resuspended in 50 mM Tris-HCl, pH 7.5, 1 mM phenylmethylsulfonyl fluoride, 1 mM EDTA and then lysed by sonication, and the cell debris was removed by centrifugation.…”

Section: Methodsmentioning

confidence: 99%

“…5A) (13). This downward curvature in the refolding arm of the chevron plot can be interpreted as arising from the population of an intermediate before the rate-limiting transition state,…”

Section: Wild Type and Mutants Refold In Low Denaturant Concentrationmentioning

confidence: 95%

“…(There is also slight curvature in the unfolding arm of the chevron plot that has been ascribed to Hammond-like movement of the transition state (13). However, the chevron plot for two-state folding, given by the solid line in Fig.…”

Section: Wild Type and Mutants Refold In Low Denaturant Concentrationmentioning

confidence: 99%

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Intermediates Control Domain Swapping during Folding of p13

Rousseau

Schymkowitz

Wilkinson

et al. 2004

Journal of Biological Chemistry

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The 13-kDa protein p13 suc1 has two folded states, a monomer and a structurally similar domain-swapped dimer formed by exchange of a ␤-strand. The refolding reaction of p13 suc1 is multiphasic, and in this paper we analyze the kinetics as a function of denaturant and protein concentration and compare the behavior of wild type and a set of mutants previously designed with dimerization propensities that span 9 orders of magnitude. We show that the folding reactions of wild type and all mutants produce the monomer predominantly despite their very different equilibrium behavior. However, the addition of low concentrations of denaturant in the refolding buffer leads to thermodynamic control of the folding reaction with products that correspond to the wild type and mutant equilibrium dimerization propensities. We present evidence that the kinetic control in the absence of urea arises because of the population of the folding intermediates. Intermediates are usually considered to be detrimental to folding because they slow down the reaction; however, our work shows that intermediates buffer the monomeric folding pathway against the effect of mutations that favor the nonfunctional, dimeric state at equilibrium. p13 suc1 (referred to subsequently as suc1), 1 a member of the Cks family of cell cycle regulatory proteins, has two native states: a monomer (1) and a domain-swapped dimer formed by exchange of a central strand, ␤4, of the ␤-sheet (2, 3) ( Fig. 1). Each subunit of the dimer is highly superimposable on the monomer except for the so-called "hinge loop" that connects the swapping strand to the rest of the structure. The hinge loop forms a ␤-hairpin in the monomer and has an extended conformation in the dimer. In this work we present an global model of the folding and interconversion of monomeric and dimeric suc1. The description illustrates how domain swapping, by exploiting mostly native interactions, creates ambiguity in protein folding that can lead to misfolding and aggregation. Unexpectedly, however, we find that intermediates can protect the folding reaction and ensure that it leads to the correct oligomeric native product.Recent theoretical and experimental work on folding of small proteins has shown that the native state topology defines the general features of protein folding pathways, and that sequence specifities modulate it to some extent (4 -6). This view raises two broad questions: 1) How do protein sequences favor native-like interactions at each stage of the folding process and avoid misfolding? Do proteins possess gatekeeper residues that restrict the conformational space to native regions (7), or is the overall energy bias in favor of native interactions rather than non-native ones sufficient to guide folding? 2) Even if only native interactions are favored, it is still possible to make native interactions with residues from another polypeptide chain rather than intramolecularly by the process of domain swapping? How do proteins avoid these undesirable interactions and ensure folding to the correct...

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Prolyl Isomerization in Protein Folding

Schmid¹

2005

Protein Folding Handbook

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Stability and folding of the cell cycle regulatory protein, p13 suc1 1 1Edited by F. E. Cohen

Cited by 33 publications

References 72 publications

Three-dimensional domain swapping in p13suc1 occurs in the unfolded state and is controlled by conserved proline residues

Three-dimensional domain swapping in p13suc1 occurs in the unfolded state and is controlled by conserved proline residues

Intermediates Control Domain Swapping during Folding of p13

Prolyl Isomerization in Protein Folding

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