Evaluation of commercially available styrene-co-maleic acid polymers for the extraction of membrane proteins from spinach chloroplast thylakoids. (May 2019)

• Record Type: Journal Article
• Authors: Korotych, Olena; Mondal, Jyotirmoy; Gattás-Asfura, Kerim M.; Hendricks, Jessica; Bruce, Barry D.
• Is Part Of: European polymer journal. Volume 114(2019)
• Page Start: 485
• Page End: 500
• Publication Date: 2019-05
• Subjects: styrene-co-maleic acid lipid particle (SMALP) -- styrene-maleic acid (SMA) copolymer -- poly(styrene-co-maleic acid) salt (pSMA-S) -- membrane protein -- thylakoid -- solubilization efficacy; APS ammonium persulfate -- ATPase adenosine triphosphatase -- ATR attenuated total reflection -- BCA bicinchoninic acid -- BSA bovine serum albumin -- CAB chlorophyll a/b binding -- CBB coomassie brilliant blue -- CN-PAGE clear native – polyacrylamide gel electrophoresis -- DMF dimethylformamide -- DDM n-dodecyl-β-D-maltoside -- DTT dithiothreitol -- EDTA ethylenediaminetetraacetic acid -- FTIR Fourier transform infrared -- IF insoluble fraction -- HRP horseradish peroxidase -- LHC light-harvesting complex -- LT low temperature -- NIST the National Institute of Standards and Technology -- PS photosystem -- pSMAnh poly(styrene-co-maleic anhydride) -- pSMA-S poly(styrene-co-maleic acid) salt -- s/m ratio styrene-to-maleic acid molar ratio -- SDS-PAGE sodium dodecyl sulfate – polyacrylamide gel electrophoresis -- SE solubilization efficacy -- SF soluble fraction -- SI supportive information -- SMA styrene-co-maleic acid -- SMALP styrene-co-maleic acid lipid particle -- TEMED tetramethylethylenediamine, N, N, N', N'- -- TM thylakoid membrane -- UV ultraviolet; Polymers -- Periodicals; Polymerization -- Periodicals; Polymères -- Périodiques; Polymérisation -- Périodiques; Polymerization; Polymers; Periodicals; Electronic journals; 547.705
• DOI: 10.1016/j.eurpolymj.2018.10.035
• Languages: English
• ISSNs: 0014-3057

Abstract

Graphical abstract: Highlights: Direct comparison of 12 commercial polymers for membrane protein (MP) extraction. Tested copolymers were both Na + and NH4 + salts of poly(styrene-co-maleic acid). The styrene-to-maleic acid molar ratio of the tested polymers ranged between 1 and 3. Only 5 polymers display high solubilization of chloroplast thylakoids. Top-5 polymers appear to yield similar qualitative results for MP extraction. Abstract: Solubilization of membrane proteins by poly(styrene-co-maleic acid) salts (pSMA-S) has significant potential for membrane protein studies. This approach provides an opportunity to overcome many disadvantages associated with a traditional detergent-based technique including protein denaturation and displacement of boundary lipids which may offer both structural and functional stability to membrane proteins. Thylakoid membranes (TMs) from photosynthetic organisms are well studied protein-rich membranes that host several multi-subunit protein complexes associated with oxygenic photosynthesis. These protein complexes are important for applied photosynthesis and by being extracted and purified they can be used in the near future for direct energy conversion. In this study, we used spinach TMs isolated from purified intact chloroplasts to systematically test the solubilization efficacy of 12 commercially available styrene-maleic acid (SMA) copolymers that vary in size, styrene-to-maleic acid molar ratio, and type of ester group. The efficacy of these pSMA-S to solubilize protein-containing biomembranes was evaluated via quantification of protein and chlorophyll content in the resulting SMA Lipid Particles (SMALPs). In addition, the extracted polymer-lipid-protein complexes were studied by low temperature fluorescence, sodium dodecyl sulfate and clear native polyacrylamide gel electrophoresis (SDS- and CN-PAGE), and immunoblot analysis. Our results indicate considerable variability in the solubilization efficacy of commercially available pSMA-S with at least 5 polymer formulations being able to efficiently extract membrane proteins from TMs. These 5 SMA copolymers may also be effective in extraction of membrane proteins from other biomembranes.