Incorporation of cytochrome b5 into supported phospholipid bilayers by vesicle fusion to supported monolayers

Kalb, Edwin; Tamm, Lukas K.

doi:10.1016/0040-6090(92)90397-t

Cited by 42 publications

(28 citation statements)

References 14 publications

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“…The question we address here is whether there is a sidedness to the deposition process. Kalb and co-workers Kalb and Tamm, 1992) have used vesicle fusion to supported monolayers to produce supported bilayers. They have proposed a mechanism that results in both the inner and outer leaflets of the vesicle forming the second monolayer.…”

Section: Resultsmentioning

confidence: 99%

Use of an oriented transmembrane protein to probe the assembly of a supported phospholipid bilayer

Contino

Hasselbacher

Ross

et al. 1994

Biophysical Journal

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Planar-supported phospholipid bilayers formed by the adsorption of vesicles are increasingly used in the investigation of lipid-dependent reactions. We have studied the way in which these bilayers are formed with phospholipid vesicles containing the transmembrane protein Tissue Factor (TF). TF complexed with the serine protease, factor VIIa, is the primary initiator of blood coagulation by way of activation of the zymogen factor X. TF has been shown to orient randomly on the inner and outer leaflets of vesicles. We used proteolytic digestion to produce vesicles in which the extracellular domain of TF is located on the inner leaflet. These vesicles show no cofactor activity for factor VIIa as a result of the inability of the extracellular domain of TF to bind VIIa. After freeze/thawing, 50% of the cofactor activity was regained, indicating reorientation of the sequestered, inner leaflet TF. Adsorption of these vesicles to the inner surface of glass microcapillaries results in a continuous phospholipid bilayer. The microcapillaries were perfused with a solution of factors VIIa and X, and the effluent was monitored for factor Xa production, a sensitive measure of the activity of the TF-VIIa complex. For coatings produced with the digested vesicles, minimal TF-VIIa activity was observed, showing that the supported bilayer preserves the orientation of the leaflets in the vesicles, i.e., the outer leaflet of the vesicles forms the outer leaflet of the supported bilayer.

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Section: Resultsmentioning

confidence: 99%

Use of an oriented transmembrane protein to probe the assembly of a supported phospholipid bilayer

Contino

Hasselbacher

Ross

et al. 1994

Biophysical Journal

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“…For integral membrane proteins, methods to incorporate the proteins into the supported planar membrane required vesicle fusion: either directly fusing vesicles that contained integral membrane proteins onto a supported substrate, such a piece of quartz or glass coverslip [45] or fusing them onto a substrate which was previously coated with a monolayer of lipids [68,69]. The mechanism of such events was not understood.…”

Section: Lipids At Interfaces: An Overviewmentioning

confidence: 99%

Bilayer-Forming Synthetic Lipids: Drugs or Carriers?

Carmona-Ribeiro

2003

CMC

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Since their introduction as bilayer-forming synthetic compounds in the eighties, dioctadecyldimethylammonium (DODA) and dihexadecylphosphate (DHP) salts have found many uses in strategic, applied areas. In particular, DODA chloride or bromide vesicles interacted with negatively charged prokaryotic or eukaryotic cells, yielding adsorption isotherms of high affinity for the cell surface, causing cell adhesion and flocculation, changing the cell surface charge from negative to positive, and causing loss of cell viability over DODA concentration ranges that depended on the cell type being tested. This work reviews data on DODA effects on cell viability (bacteria, fungus and cultured mammalian cells) to propose DODA salts as effective anti-microbial agents that exhibit differential cytotoxicity in vitro and, therefore, deserve to be investigated as potential drugs. The full utility of these inexpensive synthetic bilayers and bilayer fragments able to act as drugs themselves and, simultaneously, as drug, gene or vaccine carriers remains hitherto unexplored.

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“…Using FRAP, it was shown that the mobile fraction of the cytochrome b 5 containing SLBs is as low as 35%. This was attributed to the interaction of the protein with the underlying substrate and a possible dependence of the mobility on the orientation of the C‐terminus in the SLB . Subsequent studies utilized different substrates such as silver or gold,[29a], or biotinylated lipids in SLBs to which biotinylated cytochrome c was bound via a streptavidin linker .…”

Section: Incorporation Of Proteins Into Slbs For Light Harvesting Andmentioning

confidence: 99%

“…This was attributed to the interaction of the protein with the underlying substrate and a possible dependence of the mobility on the orientation of the C-terminus in the SLB. [ 114 ] Subsequent studies utilized different substrates such as silver or gold, [ 29a , 115 ] or biotinylated lipids in SLBs to which biotinylated cytochrome c was bound via a streptavidin linker. [ 116 ] These approaches allowed improved control over the orientation of cytochrome in SLBs.…”

Section: Cytochromesmentioning

confidence: 99%

Photosynthetic Proteins in Supported Lipid Bilayers: Towards a Biokleptic Approach for Energy Capture

Wang

Roth

Han

et al. 2015

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In nature, plants and some bacteria have evolved an ability to convert solar energy into chemical energy usable by the organism. This process involves several proteins and the creation of a chemical gradient across the cell membrane. To transfer this process in a laboratory environment several conditions have to be met: i) proteins need to be reconstituted into a lipid membrane, ii) the proteins need to be orientated and functional, and finally iii) the lipid membrane should be capable of maintaining chemical and electrical gradients. Investigating the processes of photosynthesis and energy generation in vivo is a difficult task due to the complexity of the membrane and its associated proteins. Solid supported lipid bilayers provide a good model system for systematic investigation of the different components involved in photosynthetic pathway.In this review, we describe the progress made to date in the development of supported lipid bilayer systems suitable for the investigation of membrane proteins and in particular for the reconstitution of proteins involved in light capture.

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Incorporation of cytochrome b5 into supported phospholipid bilayers by vesicle fusion to supported monolayers

Cited by 42 publications

References 14 publications

Use of an oriented transmembrane protein to probe the assembly of a supported phospholipid bilayer

Use of an oriented transmembrane protein to probe the assembly of a supported phospholipid bilayer

Bilayer-Forming Synthetic Lipids: Drugs or Carriers?

Photosynthetic Proteins in Supported Lipid Bilayers: Towards a Biokleptic Approach for Energy Capture

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