Lipophilic Fluorescent Probes: Guides to the Complexity of Lipid Membranes

Cebecauer, Marek; Šachl, Radek

doi:10.1002/9781119179320.ch16

Cited by 1 publication

(1 citation statement)

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“…A prototype of such a lipid anchored dye is obtained by the reaction of an aminereactive carboxyfluorescein derivative with phosphatidylethanolamine, first reported in 1984 by Thelen et al 135 Such lipophilic probes have been mostly used in vivo [136][137][138][139][140][141] or to characterize different membrane properties. 142 Recently, Kemmer et al have coupled the ratiometric pH sensitive dyes pHrodo or SNARF to phosphatidylethanolamine, which in contrast to fluorescein increase fluorescence upon acidification. Notably, they found a significant increase of the apparent pKa of the lipid coupled dyes in comparison to their soluble counterparts, an effect that is likely related to the close proximity of the dye to the membrane.…”

Section: Rational Design Of Fluorescent Dyes To Follow Enzyme Functionmentioning

confidence: 99%

Current problems and future avenues in proteoliposome research

Amati

Graf

Deutschmann

et al. 2020

Biochemical Society Transactions

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Membrane proteins (MPs) are the gatekeepers between different biological compartments separated by lipid bilayers. Being receptors, channels, transporters, or primary pumps, they fulfill a wide variety of cellular functions and their importance is reflected in the increasing number of drugs that target MPs. Functional studies of MPs within a native cellular context, however, is difficult due to the innate complexity of the densely packed membranes. Over the past decades, detergent-based extraction and purification of MPs and their reconstitution into lipid mimetic systems has been a very powerful tool to simplify the experimental system. In this review, we focus on proteoliposomes that have become an indispensable experimental system for enzymes with a vectorial function, including many of the here described energy transducing MPs. We first address long standing questions on the difficulty of successful reconstitution and controlled orientation of MPs into liposomes. A special emphasis is given on coreconstitution of several MPs into the same bilayer. Second, we discuss recent progress in the development of fluorescent dyes that offer sensitive detection with high temporal resolution. Finally, we briefly cover the use of giant unilamellar vesicles for the investigation of complex enzymatic cascades, a very promising experimental tool considering our increasing knowledge of the interplay of different cellular components.

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