Phosphatidylethanolamine–Lactose Permease Interaction: A Comparative Study Based on FRET

Suárez-Germà, Carme; Loura, Luís M. S.; Doménech, Òscar; Montero, M. Teresa; Vázquez-Ibar, José Luís; Hernández-Borrell, Jordi

doi:10.1021/jp309726v

Cited by 7 publications

(7 citation statements)

References 33 publications

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“…According to the data in Figure , these areas were 0.68, 0.80, and 0.90 nm 2 ·molecule –1 for DPPE, POPE, and DOPE, respectively, and 0.59, 0.68, and 0.79 nm 2 ·molecule –1 for DPPE:POPG, POPE:POPG, and DOPE:POPG mixed monolayers. The fitted values of μ show that the lipid selectivity of LacY is DOPE ∼ POPE > DPPE. − Besides, the DPPE:POPG mixture for which we have observed a double collapse in the monolayer seems not fulfill the requirements on surface lateral pressure of LacY. However, these data confirm that the molecular area occupied by a phospholipid, and hence the physical state of the phospholipid, are relevant in TMP–lipid interactions.…”

Section: Resultsmentioning

confidence: 74%

“…One of the aims of this work was to provide information on how physical properties of POPE may influence the physiological activity of LacY. As a member of the major facilitator superfamily, LacY is considered a paradigm for secondary transporters transport, the structure and function of which have been well established. , We have demonstrated previously that POPE lipid molecules are in close contact with LacY and the requirement of PEs for the correct folding of LacY in proteoliposomes has also been reported . The mechanism of action of LacY involves several steps between an initial inward-facing (∨ shape facing the periplasm) to an outward-facing (∧ shape facing the cytoplasm) conformation.…”

Section: Resultsmentioning

confidence: 99%

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Critical Temperature of 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine Monolayers and Its Possible Biological Relevance

Hernández-Borrell

Doménech

2017

J. Phys. Chem. B

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Because transmembrane proteins (TMPs) can be obtained with sufficient purity for X-ray diffraction studies more frequently than decades ago, their mechanisms of action may now be elucidated. One of the pending issues is the actual interplay between transmembrane proteins and membrane lipids. There is strong evidence of the involvement of specific lipids with some membrane proteins, such as the potassium crystallographically sited activation channel (KcsA) of Streptomyces lividans and the secondary transporter of lactose LacY of Escherichia coli, the activities of which are associated with the presence of anionic phospholipids such as the phosphatidylglycerol (PG) and phosphatidyethanolamine (PE), respectively. Other proteins such as the large conductance mechanosensitive channel (MscL) of E. coli seem to depend on the adaptation of specific phospholipids to the irregular surface of the integral membrane protein. In this work we investigated the lateral compressibility of two homoacid phosphatidylethanolamines (one with both acyl chains unsaturated (DOPE), the other with the acyl chains saturated (DPPE)) and the heteroacid phosphatidyletanolamine (POPE) and their mixtures with POPG. The liquid expanded (LE) to liquid condensed (LC) transition was observed in POPE at a temperature below its critical temperature (T = 36 °C). Because T lies below the physiological temperature, the occurrence of this phase transition may have something to do with the functioning of LacY. This magnitude is discussed within the context of the experiments carried out at temperatures below the T of POPE at which the activity of Lac Y and other TMPs are frequently studied.

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Section: Resultsmentioning

confidence: 74%

Section: Resultsmentioning

confidence: 99%

Critical Temperature of 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine Monolayers and Its Possible Biological Relevance

Hernández-Borrell

Doménech

2017

J. Phys. Chem. B

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“…These results support the stabilization role of motif A in the charge-dipole interaction. Many mutations at the +5 position of motif A in a number of MFS proteins were found to be detrimental to the transport function (33,34,36,37). For example, an Asp-to-Asn variant of Tn10 completely lost its activity (36).…”

Section: Discussionmentioning

confidence: 99%

Structure of the YajR transporter suggests a transport mechanism based on the conserved motif A

Jiang

Zhao

Wang

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

167

268

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The major facilitator superfamily (MFS) is the largest family of secondary active transporters and is present in all life kingdoms. Detailed structural basis of the substrate transport and energycoupling mechanisms of these proteins remain to be elucidated. YajR is a putative proton-driven MFS transporter found in many Gram-negative bacteria. Here we report the crystal structure of Escherichia coli YajR at 3.15 Å resolution in an outward-facing conformation. In addition to having the 12 canonical transmembrane helices, the YajR structure includes a unique 65-residue C-terminal domain which is independently stable. The structure is unique in illustrating the functional role of "sequence motif A." This highly conserved element is seen to stabilize the outward conformation of YajR and suggests a general mechanism for the conformational change between the inward and outward states of the MFS transporters.T ransporters are a type of membrane protein essential for all living cells that actively up-take nutrition and export metabolic substances and toxic materials across cellular membranes. Transporters are divided into two major types based on their energy sources. Although primary active transporters directly consume energy from ATP hydrolysis to drive substrate transport, secondary active transporters use energy derived from the electrochemical potential across the cell membrane. The major facilitator superfamily (MFS) is the largest class of secondary transporters and is present in all life kingdoms (1). For example, 25% of prokaryotic transporters belong to the MFS family (2), and the human genome contains over 110 MFS proteins (3). Currently, 3D crystal structures of nine bacterial MFS transporters (4-13) and one from fungi (9) have been reported at medium-high resolution. These studies have shown that MFS proteins contain a 12-transmembrane (TM) helix core composed of two six-helix rigid domains forming a central TM channel, which transports substrates using a rocker-switch mechanism (5). In such a mechanism, MFS proteins are believed to switch between two major conformations, inward and outward, which differ by an ∼40°rotation of one domain relative to the other. Both conformations have been captured in MFS crystal structures. However, many questions remain to be addressed, particularly those related to energy coupling and functional roles of conserved motifs.YajR, a 49-kDa transporter of the MFS family, has putatively been classified as a drug efflux protein solely on the basis of amino acid sequence analysis (14). In Escherichia coli, YajR consists of 454 amino acid residues. Besides containing 12 TM helices, YajR is predicted to possess an extra domain of about 65 residues at the C-terminal. Of the MFS proteins with reported 3D structures, the TM core of YajR shares highest sequence homology (21% identity) with EmrD (SI Appendix, Fig. S1A), which belongs to the 12-TM drug-resistance H + -driven antiporter (DHA12) subfamily (15). The YajR gene is found in a number of Gram-negative bacteria, and it shows high ...

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“…Lensink et al concluded that lipid-mediated salt-bridges are important determinants of LacY state stabilization. This mechanism of lipid-specific transporter function modulation was tested in a FRET study by Suárez-Germà et al The D/C mutation of the conserved aspartate identified by MD studies was experimentally proven to considerably change LacY selectivity for lipids. Recently, Martens et al combined hydrogen–deuterium exchange mass spectrometry (HDX-MS) with simulations and showed how the specific interactions between PE lipid headgroups and conserved charged residues (including the aspartate describe above) regulate the conformational equilibrium of the transporter .…”

Section: Slc Transportersmentioning

confidence: 99%

Emerging Diversity in Lipid–Protein Interactions

et al. 2019

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Membrane lipids interact with proteins in a variety of ways, ranging from providing a stable membrane environment for proteins to being embedded in to detailed roles in complicated and well-regulated protein functions. Experimental and computational advances are converging in a rapidly expanding research area of lipid–protein interactions. Experimentally, the database of high-resolution membrane protein structures is growing, as are capabilities to identify the complex lipid composition of different membranes, to probe the challenging time and length scales of lipid–protein interactions, and to link lipid–protein interactions to protein function in a variety of proteins. Computationally, more accurate membrane models and more powerful computers now enable a detailed look at lipid–protein interactions and increasing overlap with experimental observations for validation and joint interpretation of simulation and experiment. Here we review papers that use computational approaches to study detailed lipid–protein interactions, together with brief experimental and physiological contexts, aiming at comprehensive coverage of simulation papers in the last five years. Overall, a complex picture of lipid–protein interactions emerges, through a range of mechanisms including modulation of the physical properties of the lipid environment, detailed chemical interactions between lipids and proteins, and key functional roles of very specific lipids binding to well-defined binding sites on proteins. Computationally, despite important limitations, molecular dynamics simulations with current computer power and theoretical models are now in an excellent position to answer detailed questions about lipid–protein interactions.

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Phosphatidylethanolamine–Lactose Permease Interaction: A Comparative Study Based on FRET

Cited by 7 publications

References 33 publications

Critical Temperature of 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine Monolayers and Its Possible Biological Relevance

Critical Temperature of 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine Monolayers and Its Possible Biological Relevance

Structure of the YajR transporter suggests a transport mechanism based on the conserved motif A

Emerging Diversity in Lipid–Protein Interactions

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