Reconstitution of membrane proteins: sequential incorporation of integral membrane proteins into preformed lipid bilayers

Scotto, Anthony W.; Goodwyn, Deborah; Zakim, David

doi:10.1021/bi00377a026

Cited by 17 publications

(21 citation statements)

References 29 publications

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“…However, this was actually a membrane fusion process, rather than a transmembrane protein insertion, because bacteriorhodopsin was incorporated as small fragments of archaeal membranes containing bacteriorhodopsin already embedded in a bilayer prior to insertion. This same study [38] also reported that pig liver GDP-glucuronidyltransferase could be inserted in vesicles containing packing defects. However, this protein contains only a single C-terminal transmembrane-sequence.…”

Section: In Vitro Insertion Of Translocon-dependent Polytopic Helicalmentioning

confidence: 82%

“…For example, the archaeal proton pump bacteriorhodopsin can be inserted into unilamellar dimyristoyl phosphatidylcholine vesicles in conditions where membrane packing defects were introduced [38]. However, this was actually a membrane fusion process, rather than a transmembrane protein insertion, because bacteriorhodopsin was incorporated as small fragments of archaeal membranes containing bacteriorhodopsin already embedded in a bilayer prior to insertion.…”

Section: In Vitro Insertion Of Translocon-dependent Polytopic Helicalmentioning

confidence: 99%

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Helix insertion into bilayers and the evolution of membrane proteins

Renthal

2009

Cell. Mol. Life Sci.

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Polytopic α-helical membrane proteins cannot spontaneously insert into lipid bilayers without assistance from polytopic α-helical membrane proteins that already reside in the membrane. This raises the question of how these proteins evolved. Our current knowledge of the insertion of α-helices into natural and model membranes is reviewed with the goal of gaining insight into the evolution of membrane proteins. Topics include: translocon-dependent membrane protein insertion, antibiotic peptides and proteins, in vitro insertion of membrane proteins, chaperone-mediated insertion of transmembrane helices, and C-terminal tail-anchored (TA) proteins. Analysis of the E. coli genome reveals several predicted C-terminal TA proteins that may be descendents of proteins involved in pre-cellular membrane protein insertion. Mechanisms of pre-translocon polytopic α-helical membrane protein insertion are discussed.

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Section: In Vitro Insertion Of Translocon-dependent Polytopic Helicalmentioning

confidence: 82%

Section: In Vitro Insertion Of Translocon-dependent Polytopic Helicalmentioning

confidence: 99%

Helix insertion into bilayers and the evolution of membrane proteins

Renthal

2009

Cell. Mol. Life Sci.

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“…Free energy of incorporation depends on many parameters : water structure, protein state, lipid state, bonds between protein and water or lipid (Jahnig, 1983). It is predicted that a membrane with higher than minimal ground state energy facilitates incorporation of proteins and promotes other processes that require a modulation of bilayer organization (fusion, leakage of polar solutes, enhanced rate of transbilayer transfer of phospholipids) (Scotto and Zakim, 1985;Jaim and Zalum, 1987). The local 'organizational defects' formed then act as sites for fusion with protein aggregates.…”

Section: Discussionmentioning

confidence: 99%

Electropermeabilization mediates a stable insertion of glycophorin A with Chinese hamster ovary cell membranes

Ouagari

Benoist

Sixou

et al. 1994

European Journal of Biochemistry

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Electropulsation allowed us to incorporate glycophorin A, an integral membrane protein, into mammalian nucleated cell membranes (Chinese hamster ovary cells). The induction of stable protein association is effective only when the field intensity is higher than its threshold value, creating membrane permeabilization to small molecules. Under controlled conditions, cell viability was only slightly altered by this treatment. Pulse number and duration controlled both the number of modified cells and incorporated molecules. The phenomena was temperature dependent. An average of 5 X104 moleculeskell was bound. About 80% of cells in the pulsed population were observed to incorporate glycophorin. The protein incorporation was shown to be stable 48 h after electroassociation. Electrically bound proteins were shared between the cells after each division. As enhanced binding is detected if glycophorin is added after the pulses, it is the long-lived alteration of the membrane mediated by the pulses which supports the association.It is commonly accepted that the driving force for the spontaneous incorporation of proteins is hydrophobic effect. However, a number of other effects may support or counteract this hydrophobic effect. Generally, the free energy of protein incorporation includes four contributions which arise from a change of water structure, protein state, lipid state, bonds between protein and water or lipid molecules. Nevertheless, the central questions of how hydrophobic sequences would be transferred from an aqueous to an hydrophobic environment and how the polar regions of protein that span the membrane would be transferred across the hydrocarbon core of the bilayer, have not been emphasized. So, this makes it exceedingly unlikely that insertion of integral protein occurs directly across the lipid bilayer (Singer, 1990;Singer and Yaffe, 1990; Singer et al., 1987a, b). The membrane contribution, which has been neglected until now, necessarily appears important allowing the penetration of macromolecules with strongly polar segments. Thermodynamics of protein insertion in lipid systems imply the occurrence of the lipid matrix perturbation. Thus, protein incorporation thermodynamics ( J i m and Zakim, 1987;Jahnig, 1983) show a favourable contribution of electrostatic forces and hydrophobic forces which occur on the membrane-spanning sequence, whereas hydration forces (Pargesian and Rau, 1984) and hydrophobic forces which occur on the protein polar segments prevent incorporation. Spontaneous insertion takes place if the first component is stronger than the second one. Such a case would occur only if the membrane is in an 'excited' state.Electropulsation can create such an 'excited' state of the membrane. Recent studies reported the so-called 'electroinCorrespondence to J. Teissie,

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“…For the reconstitution of BR into egg-PC vesicles, the method based on spontaneous incorporation of integral membrane proteins into preformed lipid bilayers was used (Scotto and Zakim 1988).…”

Section: Preparation Of Hydrated Br/egg-pc Multiiayersmentioning

confidence: 99%

Influence of obstacles on lipid lateral diffusion: computer simulation of FRAP experiments and application to proteoliposomes and biomembranes

1994

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Fluorescence Recovery After Photobleaching experiments were simulated using a computer approach in which a membrane lipid leaflet was mimicked using a triangular lattice obstructed with randomly distributed immobile and non-overlapping circular obstacles. Influence of the radius r and area fraction c of these obstacles and of the radius R of the observation area on the relative diffusion coefficient D* (Eq. (1)) and mobile fraction M was analyzed. A phenomenological equation relating D* to r and c was established. Fitting this equation to the FRAP data we obtained with the probe NBD-PC embedded in bacteriorhodopsin/egg-PC multilayers suggests that this transmembrane protein rigidifies the surrounding lipid phase over a distance of about 18 A (approximately equal to two lipid layers) from the protein surface. In contrast, analysis of published diffusion constants obtained for lipids in the presence of gramicidin suggests that in terms of lateral diffusion, this relatively small polypeptide does not significantly affect the surrounding lipid phase. With respect to the mobile fraction M, and for point obstacles above the percolation threshold, an increase in R led to a decrease in M which can be associated with the existence of closed domains whose average size and diffusion properties can be determined. Adaptation of this model to the re-interpretation of the FRAP data obtained by Yechiel and Edidin (J Cell Biol (1987) 115:755-760) for the plasma membrane of human fibroblasts consistently leads to the suggestion that the lateral organization of this membrane would be of the confined type, with closed lipid domains of approximately equal to 0.5 microns 2 in area.

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Reconstitution of membrane proteins: sequential incorporation of integral membrane proteins into preformed lipid bilayers

Cited by 17 publications

References 29 publications

Helix insertion into bilayers and the evolution of membrane proteins

Helix insertion into bilayers and the evolution of membrane proteins

Electropermeabilization mediates a stable insertion of glycophorin A with Chinese hamster ovary cell membranes

Influence of obstacles on lipid lateral diffusion: computer simulation of FRAP experiments and application to proteoliposomes and biomembranes

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