How to Quantify the Chaperone-Like (Anti-Aggregation) Activity?

Kurganov, B. I.

doi:10.4172/2161-1009.1000e136

Cited by 7 publications

(6 citation statements)

References 36 publications

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“…In the case of heat-induced aggregation of γ-Cry and the DTT-induced aggregation of lysozyme, the initial parts of the kinetic curves were analyzed using quadratic equation [35][36][37]:…”

Section: Chaperone-like Activity Assaymentioning

confidence: 99%

The impact of different mutations at Arg54 on structure, chaperone-like activity and oligomerization state of human αA-crystallin: The pathomechanism underlying congenital cataract-causing mutations R54L, R54P and R54C

Khoshaman¹,

Yousefi²,

Tamaddon³

et al. 2017

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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A major part of cataractogenic mutations in human αA-Crystallin (αA-Cry) occurs at Arg residues. While Arg54 is highly conserved within different species, the cataractogenic mutations R54L, R54P and R54C have been recently identified in CRYAA gene, encoding human αA-Cry. The detailed structural and functional aspects, stability and amyloidogenic properties of αA-Cry were determined upon the above-mentioned missense mutations, using various spectroscopic techniques, gel electrophoresis, electron microscopy, size exclusion chromatography analyses, and chaperone-like activity assay. The different mutations at Arg54 result in diverse structural alterations among mutant proteins. In addition, the mutant proteins displayed reduced thermal stability, increased amyloidogenic properties and attenuated chaperone-like activity against aggregation of γ-Cry, catalase and lysozyme. The mutant proteins were also capable of forming larger oligomeric complexes with γ-Cry which is the natural partner of α-Cry in the eye lenses. The most significant structural and functional damages were observed upon R54L mutation which was also accompanied with increased oligomeric size distribution of the mutant protein. The cataractogenic nature of R54P mutation can be explained with its detrimental effect on chaperone-like activity, conformational stability and proteolytic digestibility of the mutant protein. Also, R54C αA-Cry displayed an important intrinsic propensity for disulfide protein cross-linking with significantly reduced chaperone-like activity against all client proteins. These mutations revealed a range of detrimental effects on the structure, stability and functional properties of αA-Cry which all together can explain the pathomechanisms underlying development of the associated congenital cataract disorders.

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“…In the case of heat-induced aggregation of γ-Cry and the DTT-induced aggregation of lysozyme, the initial parts of the kinetic curves were analyzed using quadratic equation [35][36][37]:…”

Section: Chaperone-like Activity Assaymentioning

confidence: 99%

The impact of different mutations at Arg54 on structure, chaperone-like activity and oligomerization state of human αA-crystallin: The pathomechanism underlying congenital cataract-causing mutations R54L, R54P and R54C

Khoshaman¹,

Yousefi²,

Tamaddon³

et al. 2017

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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show abstract

“…The kinetics of DTT-induced aggregation of α-lactalbumin was followed by dynamic light scattering (DLS). As a measure of anti-aggregation activity of α-crystallin the initial adsorption capacity of the chaperone with respect to the target protein was used [40,53,54]. The portion of α-crystallin, remaining in the native state after UV irradiation, was estimated by differential scanning calorimetry (DSC).…”

Section: Introductionmentioning

confidence: 99%

Quantification of anti-aggregation activity of UV-irradiated α-crystallin

Borzova

Markossian

Muranov

et al. 2015

International Journal of Biological Macromolecules

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“…To calculate the n value, the series of kinetic curves of inactivation at various initial protein concentrations should be obtained. The order of inactivation with respect to the protein is a power coefficient in the expression connecting the initial rate of inactivation (v 0 ) and the initial protein concentration C 0 (Borzova et al 2015;Curto et al 2012;Delahaije et al 2015;Eronina et al 2016;Kurganov 2013aKurganov , b, 2015Kurganov , 2016Le Bon et al 1999;Roefs and De Kruif 1994;van Boekel 2009):…”

Section: Dissociative Mechanism For Irreversible Thermal Denaturationmentioning

confidence: 99%

Dissociative mechanism for irreversible thermal denaturation of oligomeric proteins

2016

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Protein stability is a fundamental characteristic essential for understanding conformational transformations of the proteins in the cell. When using protein preparations in biotechnology and biomedicine, the problem of protein stability is of great importance. The kinetics of denaturation of oligomeric proteins may have characteristic properties determined by the quaternary structure. The kinetic schemes of denaturation can include the multiple stages of conformational transitions in the protein oligomer and stages of reversible dissociation of the oligomer. In this case, the shape of the kinetic curve of denaturation or the shape of the melting curve registered by differential scanning calorimetry can vary with varying the protein concentration. The experimental data illustrating dissociative mechanism for irreversible thermal denaturation of oligomeric proteins have been summarized in the present review. The use of test systems based on thermal aggregation of oligomeric proteins for screening of agents possessing anti-aggregation activity is discussed.

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How to Quantify the Chaperone-Like (Anti-Aggregation) Activity?

Cited by 7 publications

References 36 publications

The impact of different mutations at Arg54 on structure, chaperone-like activity and oligomerization state of human αA-crystallin: The pathomechanism underlying congenital cataract-causing mutations R54L, R54P and R54C

The impact of different mutations at Arg54 on structure, chaperone-like activity and oligomerization state of human αA-crystallin: The pathomechanism underlying congenital cataract-causing mutations R54L, R54P and R54C

Quantification of anti-aggregation activity of UV-irradiated α-crystallin

Dissociative mechanism for irreversible thermal denaturation of oligomeric proteins

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