The Place of Inactivated Actin and Its Kinetic Predecessor in Actin Folding−Unfolding

Kuznetsova, Irina M.; Stepanenko, Olga V.; Stepanenko, Olesya V.; Povarova, Olga I.; Biktashev, Alexander G.; Verkhusha, Vladislav V.; Shavlovsky, M. M.; Turoverov, Konstantin K.

doi:10.1021/bi026412x

Cited by 46 publications

(77 citation statements)

References 30 publications

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“…"Phase Diagram" Method of Fluorescence-The phase diagram method of fluorescence is a sensitive approach for the detection of unfolding/refolding intermediates of proteins (16,51,52). The essence of this method is to build up the diagram of I( 1 ) versus I( 2 ), where I( 1 ) and I( 2 ) are the fluorescence intensity values measured on wavelengths 1 and 2 under different experimental conditions for a protein under going structural transformations.…”

Section: Methodsmentioning

confidence: 99%

“…The essence of this method is to build up the diagram of I( 1 ) versus I( 2 ), where I( 1 ) and I( 2 ) are the fluorescence intensity values measured on wavelengths 1 and 2 under different experimental conditions for a protein under going structural transformations. Because fluorescence intensity is the extensive parameter, it will describe any two-component system by a simple relationship (16,51,52),…”

Section: Methodsmentioning

confidence: 99%

“…Moreover, each linear portion of the I( 1 ) ϭ f(I( 2 )) dependence will describe an individual all-or-none transition. In principle, 1 and 2 are arbitrary wavelengths of the fluorescence spectrum, but in practice such diagrams will be more informative if 1 and 2 will be on different slopes of the spectrum such as 320 and 365 nm (16,51,52).…”

Section: Methodsmentioning

confidence: 99%

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Unfolding of Rabbit Muscle Creatine Kinase Induced by Acid

Liang

Sanglier

et al. 2003

Journal of Biological Chemistry

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Creatine kinase (CK, 1 EC 2.7.3.2) is a key enzyme for energy homeostasis in cells and plays a significant role in the transport of high energy phosphates via phosphocreatine to sites of ATP utilization in vivo (1-4). This enzyme catalyzes the reversible phosphoryl transfer between ATP and creatine (Cr) in the presence of Mg 2ϩ , and the release of an equimolar quantity of hydrogen ion (see Reaction I).Cr ϩ MgATP 2Ϫ º PCr 2Ϫ ϩ MgADP Ϫ ϩ H ϩ REACTION I

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Unfolding of Rabbit Muscle Creatine Kinase Induced by Acid

Liang

Sanglier

et al. 2003

Journal of Biological Chemistry

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“…Phase Diagram Method of Fluorescence-The "phase diagram" method of fluorescence is a sensitive approach for the detection of unfolding/refolding intermediates of proteins (32)(33)(34)(35). The essence of this method is to build up the diagram of I( 1 ) versus I( 2 ), where I( 1 ) and I( 2 ) are the fluorescence intensity values measured on wavelengths 1 and 2 under different experimental conditions for a protein under going structural transformations.…”

Section: Determination Of Kinetic Constants For Lysozymementioning

confidence: 99%

“…In principle, 1 and 2 are arbitrary wavelengths of the fluorescence spectrum, but in practice such diagrams will be more informative if 1 and 2 are on different slopes of the spectrum, such as 320 and 365 nm (32)(33)(34)(35). It has been noted that application of this method to protein unfolding/refolding predicts that the dependence of I 320 versus I 365 will be linear if changes in the protein environment lead to an all-or-none transition between two different conformations.…”

Section: Determination Of Kinetic Constants For Lysozymementioning

confidence: 99%

Mixed Macromolecular Crowding Accelerates the Oxidative Refolding of Reduced, Denatured Lysozyme

Zhou

Liang

et al. 2004

Journal of Biological Chemistry

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The oxidative refolding of reduced, denatured hen egg white lysozyme in the presence of a mixed macromolecular crowding agent containing both bovine serum albumin (BSA) and polysaccharide has been studied from a physiological point of view. When the total concentration of the mixed crowding agent is 100 g/liter, in which the weight ratio of BSA to dextran 70 is 1:9, the refolding yield of lysozyme after refolding for 4 h under this condition increases 24% compared with that in the presence of BSA and 16% compared with dextran 70. A remarkable increase in the refolding yield of lysozyme by a mixed crowding agent containing BSA and Ficoll 70 is also observed. Further folding kinetics analyses show that these two mixed crowding agents accelerate the oxidative refolding of lysozyme remarkably, compared with single crowding agents. These results suggest that the stabilization effects of mixed macromolecular crowding agents are stronger than those of single polysaccharide crowding agents such as dextran 70 and Ficoll 70, whereas the excluded volume effects of mixed macromolecular crowding agents are weaker than those of single protein crowding agents such as BSA. Both the refolding yield and the rate of the oxidative refolding of lysozyme in these two mixed crowded solutions with suitable weight ratios are higher than those in single crowded solutions, indicating that mixed macromolecular crowding agents are more favorable to lysozyme folding and can be used to simulate the intracellular environments more accurately than single crowding agents do.The main question that protein folding encountered in vivo is that the intracellular environment is highly crowded because of the presence of high concentrations of soluble and insoluble macromolecules, which include proteins, nucleic acids, ribosomes, and carbohydrates (polysaccharides) (1-3). It has been estimated that the concentration of macromolecules in cytoplasm is in the range of 80 -400 g/liter (3-5), and all the macromolecules in physiological fluid media collectively occupy a lower limit of ϳ10% and a upper limit of ϳ40% of total fluid volume (1, 2, 6). These media are termed "crowded" or "volumeoccupied" rather than "concentrated" (7,8). The biophysical theory of macromolecular crowding has been well developed by a thermodynamic approach (2, 8 -10). The effect of excluded volume upon protein stability and conformation has been discussed using a simplified statistical thermodynamic model (11). One pertinent prediction of this model is that intramolecular excluded volume will increase the chemical potential of the unfolded state relative to that of the native or compact nonnative state, resulting in a relative stabilization of the native or compact non-native state relative to a fully unfolded state (12).Most of our knowledge about protein folding comes from biophysical studies of the refolding of pure denatured proteins at low concentrations in simple defined buffers carried out in the past three decades since the pioneering studies of Anfinsen (13). However, intra...

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Comparison of the binding behavior of FCCP with HSA and HTF as determined by spectroscopic and molecular modeling techniques

et al. 2013

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The interaction of carbonylcyanide p-(trifluoromethoxy) phenylhydrazone (FCCP) with human serum albumin (HSA) and human transferrin (HTF) was investigated using multiple spectroscopy, molecular modeling, zeta-potential and conductometry measurements of aqueous solutions at pH 7.4. The fluorescence, UV/vis and polarization fluorescence spectroscopy data disclosed that the drug-protein complex formation occurred through a remarkable static quenching. Based on the fluorescence quenching, two sets of binding sites with distinct affinities for FCCP existed in the two proteins. Steady-state and polarization fluorescence analysis showed that there were more affinities between FCCP and HSA than HTF. Far UV-CD and synchronous fluorescence studies indicated that FCCP induced more structural changes on HSA. The resonance light scattering (RLS) and zeta-potential measurements suggested that HTF had a greater resistance to drug aggregation, whereas conductometry measurements expressed the presence of free ions improving the resistance of HSA to aggregation. Thermodynamic measurements implied that a combination of electrostatic and hydrophobic forces was involved in the interaction between FCCP with both proteins. The phase diagram plots indicated that the presence of second binding site on HSA and HTF was due to the existence of intermediate structures. Site marker competitive experiments demonstrated that FCCP had two distinct binding sites in HSA which were located in sub-domains IIA and IIIA and one binding site in the C-lobe of HTF as confirmed by molecular modeling. The obtained results suggested that both proteins could act as drug carriers, but that the HSA potentially had a higher capacity for delivering FCCP to cancerous tissues.

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The Place of Inactivated Actin and Its Kinetic Predecessor in Actin Folding−Unfolding

Cited by 46 publications

References 30 publications

Unfolding of Rabbit Muscle Creatine Kinase Induced by Acid

Unfolding of Rabbit Muscle Creatine Kinase Induced by Acid

Mixed Macromolecular Crowding Accelerates the Oxidative Refolding of Reduced, Denatured Lysozyme

Comparison of the binding behavior of FCCP with HSA and HTF as determined by spectroscopic and molecular modeling techniques

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