Artificial stabilization of the fusion pore by intra-organelle styrene-maleic acid copolymers

Soft Matter ◽  
2021 ◽  
Author(s):  
Marcelo Caparotta ◽  
Marcelo Puiatti ◽  
Diego Masone
Keyword(s):  
Membrane Protein ◽  
Maleic Acid ◽  
Protein Extraction ◽  
Fusion Pore ◽  
Protein Isolation ◽  
Free Alternative ◽  
Extraction And Purification ◽  
Lipid Environment

Styrene-maleic acid copolymers have become an advantageous detergent-free alternative for membrane protein isolation. Since their discovery, experimental membrane protein extraction and purification by keeping intact their lipid environment has become...

scholarly journals Membrane protein extraction and purification using partially-esterified SMA polymers

2021 ◽  
pp. 183758
Author(s):  
Olivia P. Hawkins ◽  
Christine Parisa T. Jahromi ◽  
Aiman A. Gulamhussein ◽  
Stephanie Nestorow ◽  
Taranpreet Bahra ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Protein Extraction ◽  
Extraction And Purification

scholarly journals Membrane protein extraction and purification using styrene–maleic acid (SMA) copolymer: effect of variations in polymer structure

2016 ◽  
Vol 473 (23) ◽  
pp. 4349-4360 ◽  
Author(s):  
Kerrie A. Morrison ◽  
Aneel Akram ◽  
Ashlyn Mathews ◽  
Zoeya A. Khan ◽  
Jaimin H. Patel ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Membrane Protein ◽  
Maleic Acid ◽  
Protein Extraction ◽  
Affinity Purification ◽  
Expression System ◽  
Transmembrane Proteins ◽  
Membrane Extraction ◽  
Total Size ◽  
Average Molecular Mass

The use of styrene–maleic acid (SMA) copolymers to extract and purify transmembrane proteins, while retaining their native bilayer environment, overcomes many of the disadvantages associated with conventional detergent-based procedures. This approach has huge potential for the future of membrane protein structural and functional studies. In this investigation, we have systematically tested a range of commercially available SMA polymers, varying in both the ratio of styrene and maleic acid and in total size, for the ability to extract, purify and stabilise transmembrane proteins. Three different membrane proteins (BmrA, LeuT and ZipA), which vary in size and shape, were used. Our results show that several polymers, can be used to extract membrane proteins, comparably to conventional detergents. A styrene:maleic acid ratio of either 2:1 or 3:1, combined with a relatively small average molecular mass (7.5–10 kDa), is optimal for membrane extraction, and this appears to be independent of the protein size, shape or expression system. A subset of polymers were taken forward for purification, functional and stability tests. Following a one-step affinity purification, SMA 2000 was found to be the best choice for yield, purity and function. However, the other polymers offer subtle differences in size and sensitivity to divalent cations that may be useful for a variety of downstream applications.

scholarly journals Membrane Protein Extraction by Styrene-Maleic Acid Copolymers is Independent of Subcellular Localization but is Influenced by Membrane Organization

2017 ◽  
Vol 112 (3) ◽  
pp. 385a
Author(s):  
Jonas M. Doerr ◽  
Juan J. Dominguez Pardo ◽  
Marleen H. van Coevoorden-Hameete ◽  
Casper C. Hoogenraad ◽  
J. Antoinette Killian
Keyword(s):  
Membrane Protein ◽  
Subcellular Localization ◽  
Maleic Acid ◽  
Protein Extraction ◽  
Membrane Organization

Protein Extraction Efficiency and Selectivity of Esterified Styrene–Maleic Acid Copolymers in Thylakoid Membranes

2021 ◽  
Author(s):  
Nathan G. Brady ◽  
Cameron E. Workman ◽  
Bridgie Cawthon ◽  
Barry D. Bruce ◽  
Brian K. Long
Keyword(s):  
Maleic Acid ◽  
Extraction Efficiency ◽  
Protein Extraction ◽  
Thylakoid Membranes

Order-disorder transitions of cytoplasmic N-termini in the mechanisms of P-type ATPases

2020 ◽  
Author(s):  
Khondker Rufaka Hossain ◽  
Daniel Clayton ◽  
Sophia C Goodchild ◽  
Alison Rodger ◽  
Richard James Payne ◽  
...  
Keyword(s):  
Protein Structure ◽  
Membrane Protein ◽  
Structure And Function ◽  
Membrane Protein Structure ◽  
Protein Structure And Function ◽  
Lipid Environment ◽  
Membrane Pumps ◽  
And Function ◽  
P Type

Membrane protein structure and function are modulated via interactions with their lipid environment. This is particularly true for the integral membrane pumps, the P-type ATPases. These ATPases play vital roles...

Functional Integrity of Membrane Protein Rhodopsin Solubilized by Styrene-Maleic Acid Copolymer

2021 ◽  
Author(s):  
Stephanie G. Pitch ◽  
Weekie Yao ◽  
Istvan Szundi ◽  
Jonathan Fay ◽  
Eefei Chen ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Maleic Acid ◽  
Functional Integrity ◽  
Acid Copolymer ◽  
Maleic Acid Copolymer

scholarly journals Selective Enrichment of Membrane Proteins by Partition Phase Separation for Proteomic Studies

2003 ◽  
Vol 2003 (4) ◽  
pp. 249-255 ◽  
Author(s):  
M. Walid Qoronfleh ◽  
Betsy Benton ◽  
Ray Ignacio ◽  
Barbara Kaboord
Keyword(s):  
Membrane Proteins ◽  
Membrane Protein ◽  
Protein Extraction ◽  
Yeast Cells ◽  
Protein Markers ◽  
Two Phase ◽  
Western Blots ◽  
Selective Enrichment ◽  
Phase Partitioning ◽  
Hydrophobic Fraction

The human proteome project will demand faster, easier, and more reliable methods to isolate and purify protein targets. Membrane proteins are the most valuable group of proteins since they are the target for 70–80% of all drugs. Perbio Science has developed a protocol for the quick, easy, and reproducible isolation of integral membrane proteins from eukaryotic cells. This procedure utilizes a proprietary formulation to facilitate cell membrane disruption in a mild, nondenaturing environment and efficiently solubilizes membrane proteins. The technique utilizes a two-phase partitioning system that enables the class separation of hydrophobic and hydrophilic proteins. A variety of protein markers were used to investigate the partitioning efficiency of the membrane protein extraction reagents (Mem-PER) (Mem-PER is a registered trademark of Pierce Biotechnology, Inc) system. These included membrane proteins with one or more transmembrane spanning domains as well as peripheral and cytosolic proteins. Based on densitometry analyses of our Western blots, we obtained excellent solubilization of membrane proteins with less than 10% contamination of the hydrophobic fraction with hydrophilic proteins. Compared to other methodologies for membrane protein solubilization that use time-consuming protocols or expensive and cumbersome instrumentation, the Mem-PER reagents system for eukaryotic membrane protein extraction offers an easy, efficient, and reproducible method to isolate membrane proteins from mammalian and yeast cells.

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