The influence of organic and inorganic additives on the polymer mediated phase separation of Triton X-100

2021 ◽  
pp. 116182
Author(s):  
Md. Monir Hosen ◽  
Sharmin Sultana Rakhi ◽  
M. Alfakeer ◽  
Mohammad Majibur Rahman ◽  
Shamim Mahbub ◽  
...  
Keyword(s):  
Phase Separation ◽  
Triton X

scholarly journals Phase separation of the receptor for immunoglobulin E and its subunits in Triton X-114.

1984 ◽  
Vol 259 (23) ◽  
pp. 14922-14927 ◽  
Author(s):  
G Alcaraz ◽  
J P Kinet ◽  
N Kumar ◽  
S A Wank ◽  
H Metzger
Keyword(s):  
Phase Separation ◽  
Immunoglobulin E ◽  
Triton X

scholarly journals Glycosyl-phosphatidylinositol-anchored membrane proteins can be distinguished from transmembrane polypeptide-anchored proteins by differential solubilization and temperature-induced phase separation in Triton X-114

1991 ◽  
Vol 280 (3) ◽  
pp. 745-751 ◽  
Author(s):  
N M Hooper ◽  
A Bashir
Keyword(s):  
Phase Separation ◽  
Membrane Proteins ◽  
Aqueous Phase ◽  
Hydrophobic Membrane ◽  
Rich Phase ◽  
Glycosyl Phosphatidylinositol ◽  
Ionic Detergent ◽  
Membrane Spanning ◽  
Triton X ◽  
Insoluble Pellet

Treatment of kidney microvillar membranes with the non-ionic detergent Triton X-114 at 0 degrees C, followed by low-speed centrifugation, generated a detergent-insoluble pellet and a detergent-soluble supernatant. The supernatant was further fractionated by phase separation at 30 degrees C into a detergent-rich phase and a detergent-depleted or aqueous phase. Those ectoenzymes with a covalently attached glycosyl-phosphatidylinositol (G-PI) membrane anchor were recovered predominantly (greater than 73%) in the detergent-insoluble pellet. In contrast, those ectoenzymes anchored by a single membrane-spanning polypeptide were recovered predominantly (greater than 62%) in the detergent-rich phase. Removal of the hydrophobic membrane-anchoring domain from either class of ectoenzyme resulted in the proteins being recovered predominantly (greater than 70%) in the aqueous phase. This technique was also applied to other membrane types, including pig and human erythrocyte ghosts, where, in both cases, the G-PI-anchored acetylcholinesterase partitioned predominantly (greater than 69%) into the detergent-insoluble pellet. When the microvillar membranes were subjected only to differential solubilization with Triton X-114 at 0 degrees C, the G-PI-anchored ectoenzymes were recovered predominantly (greater than 63%) in the detergent-insoluble pellet, whereas the transmembrane-polypeptide-anchored ectoenzymes were recovered predominantly (greater than 95%) in the detergent-solubilized supernatant. Thus differential solubilization and temperature-induced phase separation in Triton X-114 distinguished between G-PI-anchored membrane proteins, transmembrane-polypeptide-anchored proteins and soluble, hydrophilic proteins. This technique may be more useful and reliable than susceptibility to release by phospholipases as a means of identifying a G-PI anchor on an unpurified membrane protein.

scholarly journals The hyaluronate synthase from a eukaryotic cell line

1993 ◽  
Vol 290 (3) ◽  
pp. 791-795 ◽  
Author(s):  
L Klewes ◽  
E A Turley ◽  
P Prehm
Keyword(s):  
Monoclonal Antibodies ◽  
Phase Separation ◽  
Affinity Chromatography ◽  
Cell Line ◽  
Aqueous Phase ◽  
Plasma Membranes ◽  
Eukaryotic Cell ◽  
Molecular Masses ◽  
Triton X ◽  
Hyaluronate Synthase

The hyaluronate synthase complex was identified in plasma membranes from B6 cells. It contained two subunits of molecular masses 52 kDa and 60 kDa which bound the precursor UDP-GlcA in digitonin solution and partitioned into the aqueous phase, together with nascent hyaluronate upon Triton X-114 phase separation. The 52 kDa protein cross-reacted with poly- and monoclonal antibodies raised against the streptococcal hyaluronate synthase and the 60 kDa protein was recognized by monoclonal antibodies raised against a hyaluronate receptor. The 52 kDa protein was purified to homogeneity by affinity chromatography with monoclonal anti-hyaluronate synthase.

Membrane association and isolation of the S-layer protein of Methanoculleus marisnigri

1994 ◽  
Vol 40 (3) ◽  
pp. 237-241 ◽  
Author(s):  
Douglas P. Bayley ◽  
Susan F. Koval
Keyword(s):  
Plasma Membrane ◽  
Phase Separation ◽  
Lattice Spacing ◽  
Guanidine Hydrochloride ◽  
Membrane Association ◽  
Soluble Extract ◽  
Membrane Complex ◽  
Layer Membrane ◽  
Triton X ◽  
Anaerobic Medium

Methanoculleus marisnigri is an irregularly shaped coccoid member of the methanogenic archaeobacteria. The cells possessed a hexagonally arranged, glycosylated S-layer as the sole wall component. The lattice spacing was approximately 13.5 nm. Plasmolysis in anaerobic medium with 20% sucrose did not separate the plasma membrane from the S-layer. The S-layer–membrane complex formed a tight but noncovalent association, was deformable, and was not a rigid structure. The 138-kDa glycoprotein was not solubilized by guanidine hydrochloride or urea, but it was solubilized in the detergent Triton X-100 at temperatures above 60 °C, and purified by phase separation at 75 °C of the detergent-soluble extract. The amino acid composition of the glycoprotein was similar to that reported for other S-layer proteins.Key words: methanogen, archaeobacteria (archaea), S-layer, glycoprotein, Triton X-100.

scholarly journals Acetylcholinesterase from Apis mellifera head. Evidence for amphiphilic and hydrophilic forms characterized by Triton X-114 phase separation

1988 ◽  
Vol 255 (2) ◽  
pp. 463-470 ◽  
Author(s):  
L P Belzunces ◽  
J P Toutant ◽  
M Bounias
Keyword(s):  
Phase Separation ◽  
Apis Mellifera ◽  
Aqueous Phase ◽  
Sucrose Gradient ◽  
Electrophoretic Analysis ◽  
High Salt ◽  
Sedimentation Analysis ◽  
Low Salt ◽  
S Peak ◽  
Triton X

The polymorphism of bee acetylcholinesterase was studied by sucrose-gradient-sedimentation analysis and non-denaturing electrophoretic analysis of fresh extracts. Lubrol-containing extracts exhibited only one form, which sedimented at 5 S when analysed on high-salt Lubrol-containing gradients and 6 S when analysed on low-salt Lubrol-containing gradients. The 5 S/6 S form aggregated upon removal of the detergent when sedimented on detergent-free gradients and was recovered in the detergent phase after Triton X-114 phase separation. Thus the 5 S/6 S enzyme corresponds to an amphiphilic acetylcholinesterase form. In detergent-free extracts three forms, whose apparent sedimentation coefficients are 14 S, 11 S and 7 S, were observed when sedimentations were performed on detergent-free gradients. Sedimentation analyses on detergent-containing gradients showed only a 5 S peak in high-salt detergent-free extracts and a 6 S peak, with a shoulder at about 7 S, in low-salt detergent-free extracts. Electrophoretic analysis in the presence of detergent demonstrated that the 14 S and 11 S peaks corresponded to aggregates of the 5 S/6 S form, whereas the 7 S peak corresponded to a hydrophilic acetylcholinesterase form which was recovered in the aqueous phase following Triton X-114 phase separation. The 5 S/6 S amphiphilic form could be converted into a 7.1 S hydrophilic form by phosphatidylinositol-specific phospholipase C digestion.

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