Effect of crowding on several stages of protein aggregation in test systems in the presence of α-crystallin

Chebotareva, Natalia A.; Filippov, Dmitrii O.; Bi, Kurganov

doi:10.1016/j.ijbiomac.2015.07.002

Cited by 22 publications

(13 citation statements)

References 56 publications

(76 reference statements)

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“…On the basis of analysis of the dependence of parameter K LS on the protein concentration the conclusion has been made that DTTinduced aggregation of α-lactalbumin proceeds in the kinetic regime where the rate-limiting stage is the stage of aggregation of unfolded protein molecules. The value of n for the aggregation process was found to be 2.5 ± 0.2 (80 mM Hepes buffer, рН 6.8, containing 0.1 M NaCl; 37°C) [17]. The data obtained in our earlier works are consistent with this conclusion [10,18,19].…”

Section: Editorialsupporting

confidence: 83%

Selection of Test Systems for Estimation of Anti-aggregation Activity of Molecular Chaperones

Kurganov¹

2015

Biochem Anal Biochem

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Section: Editorialsupporting

confidence: 83%

Selection of Test Systems for Estimation of Anti-aggregation Activity of Molecular Chaperones

Kurganov¹

2015

Biochem Anal Biochem

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“…These environmental perturbations significantly influence a variety of biologically-relevant processes, such as the folding process, the binding of small molecules, enzymatic activity, protein-protein interactions, amyloid formation, etc. [24,27,28,29,30,31,32]. In laboratory practice, different polymer molecules, such as polyethylene glycol (PEG) of different molecular weights, Dextran, Ficoll, and inert proteins, are typically used as crowding agents [25,28,29,31].…”

Section: Introductionmentioning

confidence: 99%

Peculiarities of the Super-Folder GFP Folding in a Crowded Milieu

Stepanenko

Kuznetsova

Uversky

et al. 2016

IJMS

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The natural cellular milieu is crowded by large quantities of various biological macromolecules. This complex environment is characterized by a limited amount of unoccupied space, limited amounts of free water, and changed solvent properties. Obviously, such a tightly packed cellular environment is poorly mimicked by traditional physiological conditions, where low concentrations of a protein of interest are analyzed in slightly salted aqueous solutions. An alternative is given by the use of a model crowded milieu, where a protein of interest is immersed in a solution containing high concentrations of various polymers that serve as model crowding agents. An expected outcome of the presence of such macromolecular crowding agents is their ability to increase conformational stability of a globular protein due to the excluded volume effects. In line with this hypothesis, the behavior of a query protein should be affected by the hydrodynamic size and concentration of an inert crowder (i.e., an agent that does not interact with the protein), whereas the chemical nature of a macromolecular crowder should not play a role in its ability to modulate conformational properties. In this study, the effects of different crowding agents (polyethylene glycols (PEGs) of various molecular masses (PEG-600, PEG-8000, and PEG-12000), Dextran-70, and Ficoll-70) on the spectral properties and unfolding–refolding processes of the super-folder green fluorescent protein (sfGFP) were investigated. sfGFP is differently affected by different crowders, suggesting that, in addition to the expected excluded volume effects, there are some changes in the solvent properties.

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“…Studies in this field revealed that the efficiency of crowding agents might depend on the ratio between the hydrodynamic dimensions (or occupied volumes) of the crowder and the test molecule, with the most effective conditions being those where the crowder and the test molecule occupy similar volumes ( Chen et al, 2011 ; Minton, 1993 ; Tokuriki et al, 2004 ). Typically, high concentrations of inert crowders have significant effects on conformational stability and structural properties of some proteins ( Christiansen et al, 2010 ; Engel et al, 2008 ; Kuznetsova, Turoverov & Uversky, 2014 ; Mittal & Singh, 2013 ), and may affect various biological processes, such as protein folding, binding of small molecules, enzymatic activity, protein-nucleic acid interactions, protein-protein interactions, protein chaperone activity, pathological protein aggregation, and extent of amyloid formation ( Chebotareva et al, 2015a ; Chebotareva, Filippov & Kurganov, 2015b ; Hatters, Minton & Howlett, 2002 ; Kuznetsova, Turoverov & Uversky, 2014 ; Kuznetsova et al, 2015 ; Minton, 2000a ; Morar et al, 2001 ; Shtilerman, Ding & Lansbury, 2002 ; Uversky et al, 2002 ). For example, we recently conducted a large-scale analysis of the effect of two traditional macromolecular crowders, PEG-8000 and Dextran-70, on the urea-induced unfolding of eleven globular proteins belonging to different structural classes ( Stepanenko et al, 2016a ).…”

Section: Introductionmentioning

confidence: 99%

Structure and stability of recombinant bovine odorant-binding protein: III. Peculiarities of the wild type bOBP unfolding in crowded milieu

Kuznetsova

et al. 2016

PeerJ

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Contrary to the majority of the members of the lipocalin family, which are stable monomers with the specific OBP fold (a β-barrel consisting of a 8-stranded anti-parallel β-sheet followed by a short α-helical segment, a ninth β-strand, and a disordered C-terminal tail) and a conserved disulfide bond, bovine odorant-binding protein (bOBP) does not have such a disulfide bond and forms a domain-swapped dimer that involves crossing the α-helical region from each monomer over the β-barrel of the other monomer. Furthermore, although natural bOBP isolated from bovine tissues exists as a stable domain-swapped dimer, recombinant bOBP has decreased dimerization potential and therefore exists as a mixture of monomeric and dimeric variants. In this article, we investigated the effect model crowding agents of similar chemical nature but different molecular mass on conformational stability of the recombinant bOBP. These experiments were conducted in order to shed light on the potential influence of model crowded environment on the unfolding-refolding equilibrium. To this end, we looked at the influence of PEG-600, PEG-4000, and PEG-12000 in concentrations of 80, 150, and 300 mg/mL on the equilibrium unfolding and refolding transitions induced in the recombinant bOBP by guanidine hydrochloride. We are showing here that the effect of crowding agents on the structure and conformational stability of the recombinant bOBP depends on the size of the crowder, with the smaller crowding agents being more effective in the stabilization of the bOBP native dimeric state against the guanidine hydrochloride denaturing action. This effect of the crowding agents is concentration dependent, with the high concentrations of the agents being more effective.

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Effect of crowding on several stages of protein aggregation in test systems in the presence of α-crystallin

Cited by 22 publications

References 56 publications

Selection of Test Systems for Estimation of Anti-aggregation Activity of Molecular Chaperones

Selection of Test Systems for Estimation of Anti-aggregation Activity of Molecular Chaperones

Peculiarities of the Super-Folder GFP Folding in a Crowded Milieu

Structure and stability of recombinant bovine odorant-binding protein: III. Peculiarities of the wild type bOBP unfolding in crowded milieu

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