Detergent-Free Isolation, Characterization and Functional Reconstitution of a K+ Channel: The Power of Native Nanodiscs

Dörr, Jonas M.; Schäfer, Marre; Koorengevel, Martijn C.; Killian, J. Antoinette

doi:10.1016/j.bpj.2015.11.2274

Cited by 1 publication

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“…Native nanodiscs typically have a diameter ranging from 10 to 24 nm, while the SMA copolymer has a thickness of approximately 1 nm [ 81 ]. Recently, this approach has been successfully used to isolate various membrane-associated proteins, such as proton pumps, photosynthetic systems, and ion channels [ 79 , 82 , 83 ]. Consistently, the transfer of an ion channel, isolated in native nanodiscs, to a planar membrane resulted in a transmembrane ion conductance [ 84 ].…”

Section: Detergent-free Native-like Membrane Proteins’ Reconstitution...mentioning

confidence: 99%

Reviving Natural Rubber Synthesis via Native/Large Nanodiscs

Umar,

Ahmad,

2024

Polymers

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Natural rubber (NR) is utilized in more than 40,000 products, and the demand for NR is projected to reach $68.5 billion by 2026. The primary commercial source of NR is the latex of Hevea brasiliensis. NR is produced by the sequential cis-condensation of isopentenyl diphosphate (IPP) through a complex known as the rubber transferase (RTase) complex. This complex is associated with rubber particles, specialized organelles for NR synthesis. Despite numerous attempts to isolate, characterize, and study the RTase complex, definitive results have not yet been achieved. This review proposes an innovative approach to overcome this longstanding challenge. The suggested method involves isolating the RTase complex without using detergents, instead utilizing the native membrane lipids, referred to as “natural nanodiscs”, and subsequently reconstituting the complex on liposomes. Additionally, we recommend the adaptation of large nanodiscs for the incorporation and reconstitution of the RTase complex, whether it is in vitro transcribed or present within the natural nanodiscs. These techniques show promise as a viable solution to the current obstacles. Based on our experimental experience and insights from published literature, we believe these refined methodologies can significantly enhance our understanding of the RTase complex and its role in in vitro NR synthesis.

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