Thermal Denaturation and Aggregation Assays in Analytical Biochemistry

Kurganov, B. I.

doi:10.4172/2161-1009.1000e143

Cited by 8 publications

(2 citation statements)

References 53 publications

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“…It fails to serve as a reporter for FSEC-TS of membrane proteins with apparent T m exceeding the T m of GFP itself which is~76°C (10-min heating) 46 . While this appears to be reasonably high, it is important to note that the actual T m may be far below the apparent T m (or more appropriately termed as T aggregation ) because proteins can undergo denaturation long before aggregation 37,55 . Therefore, sometimes conditions leading to apparent T m of higher-than-76°C need to be sought.…”

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confidence: 95%

An improved fluorescent tag and its nanobodies for membrane protein expression, stability assay, and purification

Cai

Yao

et al. 2020

Commun Biol

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Green fluorescent proteins (GFPs) are widely used to monitor membrane protein expression, purification, and stability. An ideal reporter should be stable itself and provide high sensitivity and yield. Here, we demonstrate that a coral (Galaxea fascicularis) thermostable GFP (TGP) is by such reasons an improved tag compared to the conventional jellyfish GFPs. TGP faithfully reports membrane protein stability at temperatures near 90 °C (20-min heating). By contrast, the limit for the two popular GFPs is 64 °C and 74 °C. Replacing GFPs with TGP increases yield for all four test membrane proteins in four expression systems. To establish TGP as an affinity tag for membrane protein purification, several high-affinity synthetic nanobodies (sybodies), including a non-competing pair, are generated, and the crystal structure of one complex is solved. Given these advantages, we anticipate that TGP becomes a widely used tool for membrane protein structural studies.

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confidence: 95%

An improved fluorescent tag and its nanobodies for membrane protein expression, stability assay, and purification

Cai

Yao

et al. 2020

Commun Biol

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“…To interpret the effects of different agents on aggregation of target proteins, the additional experiments on protein stability in the presence of the agents under study should be carried out. It is worth noting that test systems of type A can be used for screening of ligands specifically interacting with proteins on the basis of measurements of the aggregation rate in the presence of ligands under study [23][24][25][26].…”

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confidence: 99%

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

Kurganov¹

2015

Biochem Anal Biochem

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Relationship between the initial rate of protein aggregation and the lag period for amorphous aggregation

Borzova

Markossian

2014

International Journal of Biological Macromolecules

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Thermal Denaturation and Aggregation Assays in Analytical Biochemistry

Cited by 8 publications

References 53 publications

An improved fluorescent tag and its nanobodies for membrane protein expression, stability assay, and purification

An improved fluorescent tag and its nanobodies for membrane protein expression, stability assay, and purification

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

Relationship between the initial rate of protein aggregation and the lag period for amorphous aggregation

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