Evaluation of the surface hydrophobicities of proteins using aqueous two-phase partitioning systems and application to the separation process.

Yano, Koji; Wakayama, Akihiro; Kuboi, Ryoichi; Komasawa, Isao

doi:10.2116/bunsekikagaku.42.11_673

Cited by 10 publications

(3 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A quantitative evaluation of the local surface hydrophobicity of proteins has been conducted by using APTPS containing Triton X-405. 123,124 When Triton X-405 is added to the Dx-PEG system, the proteins having hydrophobic sites interact with Triton X-405 and tend to partition into the PEG-rich phase. Thus, the local hydrophobicity of the protein can be quantified from the change in the partition coefficient of the protein with increase in the Triton X-405 concen-tration.…”

Section: •6 Aptps Including Poeae Surfactantsmentioning

confidence: 99%

Aqueous Micellar Two-Phase Systems for Protein Separation

1998

View full text Add to dashboard Cite

The extractive technique for protein purification based on two-phase separation in aqueous micellar solutions (aqueous micellar two-phase system (AMTPS)) is reviewed. The micellar solution of a nonionic surfactant, such as polyoxyethylene alkyl ether, which is most frequently used for protein extraction, separates into two phases upon heating above its cloud point. The two phases consist of a surfactant-depleted phase (aqueous phase) and a surfactant-rich phase. Hydrophilic proteins are partitioned to the aqueous phase and hydrophobic membrane proteins are extracted into the surfactant-rich phase. Because of the methodological simplicity and rapidity, this technique has become an effective means, and thus has been widely used for the purification and characterization of proteins. In contrast to polyoxyethylene alkyl ether, micellar solutions of a zwitterionic surfactant, such as alkylammoniopropyl sulfate, separate below the critical temperature. Alkylglucosides can also separate into two phases upon adding water-soluble polymers. Recently, these twophase systems have been exploited for protein separation. Additionally, hydrophobic affinity ligands, charged polymers, and ionic surfactants have been successfully used for controlling the extractability of proteins in AMTPS.

show abstract

Section: •6 Aptps Including Poeae Surfactantsmentioning

confidence: 99%

Aqueous Micellar Two-Phase Systems for Protein Separation

1998

View full text Add to dashboard Cite

show abstract

“…In general, sufficient CCC separation between components 1 and 2 could be achieved if the separation factor (K 2 /K 1 ) is more than 1.5. In the aqueous two-phase solvent system, the K value of protein is calculated using the following formula (Yano et al, 1993):…”

Section: Future Perspective Of CCCmentioning

confidence: 99%

“…In general, sufficient CCC separation between components 1 and 2 could be achieved if the separation factor ( K 2 / K 1 ) is more than 1.5. In the aqueous two‐phase solvent system, the K value of protein is calculated using the following formula (Yano et al, 1993):

\ln K goodbreak= \ln K_{el} goodbreak+ \ln K_{hphob}

where K el represents the contribution by electrostatic effect that can be neglected at the isoelectric point (p I ) and K hphob represents the contribution by hydrophobic effect that shows the surface hydrophobicity of the analyte protein. Most proteins are charged in the aqueous solution so that they partition into a more hydrophilic layer in the two‐phase solvent system.…”

Section: Future Perspective Of CCCmentioning

confidence: 99%

Countercurrent chromatography as a frontier tool to discover new mechanical roles in liquid–liquid phase separation of cellular biomolecules

Shinomiya

Iijima

Ito

2023

Biomedical Chromatography

View full text Add to dashboard Cite

The development of countercurrent chromatography (CCC) technology enabled us to achieve higher peak resolutions and more shortened separation times even for protein separation using aqueous two-phase solvent systems composed of polyethylene glycol and inorganic salts (or dextrans). By eliminating the solid support matrix, all analytes can be recovered from the coiled column after the separation is completed.Recently, it has been found that droplets of biomolecules formed by liquid-liquid phase separation in cells closely relate to the transcription, regulation of signal transduction, and formation of amyloids. Meanwhile, although CCC is a separation technique based on liquid-liquid partitioning of analytes between two immiscible phases, the mechanism of separation could suggest some idea concerning the formation of biomolecule droplets in cells. This article describes the recent advances in the CCC apparatus, the coiled separation column, the choice of a suitable two-phase solvent system, and the application to separation and purification of bioactive macromolecules such as proteins and enzymes, and also discusses the possibility of CCC as a tool to reveal new mechanical roles of biomolecule droplets in the cellular environments.aqueous two-phase solvent system, cellular biomolecules, coil planet centrifuge, coil satellite centrifuge, countercurrent chromatography 1 | INTRODUCTION Recently, it has been found that some proteins in cells function as aggregates after forming droplets in the liquid-liquid phase separation (LLPS) process. This discovery closely relates to various biological behaviors, including transcription, regulation of signal transduction, and formation of amyloids. The transcription factor forms droplets congregated by an electrostatic interaction between the intrinsically disordered domain and DNA and transferred into a higher state of transcription activity (Chong et al., 2018). The study on signal transduction mechanism shows that biomacromolecules connect with one another by weak interaction, and in proteins the intrinsically disordered regions generally relate to their interactions. Stress granules formed temporally in cytoplasm regulate stress response signals by holding various transduction molecules in themselves (Riggs et al., 2020). Furthermore, amyloid, protein aggregate often formed by the failure of folding, is attributed to serious diseases, that is, neurodegenerative diseases such as Alzheimer's disease, Huntington's

show abstract