Membrane Protein Production in Lactococcus lactis for Functional Studies

Seigneurin-Berny, Daphné; King, Martin S.; Sautron, Emiline; Moyet, Lucas; Catty, Patrice; André, François; Rolland, Norbert; Kunji, Edmund R. S.; Frelet-Barrand, Annie

doi:10.1007/978-1-4939-3637-3_6

Cited by 2 publications

(2 citation statements)

References 48 publications

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“…As far as we know, the description of ectopic intracellular membrane generation in other bacteria than E. coli has not been reported yet. The Gram-positive bacteria Lactococcus lactis is well known to allow for high-level production of exogenous membrane proteins without forming inclusion bodies [ 107 ]. Therefore, the formation of internal neo-membranes within these bacteria could be a convenient means for resisting against membrane stress and cell toxicity that is induced by the accumulation of the neo-synthesized proteins, but this has not been addressed yet.…”

Section: Some Remaining Questionsmentioning

confidence: 99%

Ectopic Neo-Formed Intracellular Membranes in Escherichia coli: A Response to Membrane Protein-Induced Stress Involving Membrane Curvature and Domains

et al. 2018

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Bacterial cytoplasmic membrane stress induced by the overexpression of membrane proteins at high levels can lead to formation of ectopic intracellular membranes. In this review, we report the various observations of such membranes in Escherichia coli, compare their morphological and biochemical characterizations, and we analyze the underlying molecular processes leading to their formation. Actually, these membranes display either vesicular or tubular structures, are separated or connected to the cytoplasmic membrane, present mono- or polydispersed sizes and shapes, and possess ordered or disordered arrangements. Moreover, their composition differs from that of the cytoplasmic membrane, with high amounts of the overexpressed membrane protein and altered lipid-to-protein ratio and cardiolipin content. These data reveal the importance of membrane domains, based on local specific lipid–protein and protein–protein interactions, with both being crucial for local membrane curvature generation, and they highlight the strong influence of protein structure. Indeed, whether the cylindrically or spherically curvature-active proteins are actively curvogenic or passively curvophilic, the underlying molecular scenarios are different and can be correlated with the morphological features of the neo-formed internal membranes. Delineating these molecular mechanisms is highly desirable for a better understanding of protein–lipid interactions within membrane domains, and for optimization of high-level membrane protein production in E. coli.

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Section: Some Remaining Questionsmentioning

confidence: 99%

Ectopic Neo-Formed Intracellular Membranes in Escherichia coli: A Response to Membrane Protein-Induced Stress Involving Membrane Curvature and Domains

et al. 2018

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“…There are many potential issues with their expression, targeting, insertion and folding, when they are produced through heterologous expression. In the Gram-negative bacterium Escherichia coli they end up in inclusion bodies in a misfolded and aggregated state [26], whereas in the Gram-positive bacterium Lactococcus lactis they are targeted to the cytoplasmic membrane [27][28][29][30], but a fraction might be misfolded [28]. Mitochondrial carriers expressed in the inner mitochondrial membrane of yeast or mammalian cells, using the endogenous synthesis, targeting, insertion and folding pathways, are the best starting point.…”

Section: Introductionmentioning

confidence: 99%

The mitochondrial ADP/ATP carrier exists and functions as a monomer

Kunji

Ruprecht

2020

Biochemical Society Transactions

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For more than 40 years, the oligomeric state of members of the mitochondrial carrier family (SLC25) has been the subject of debate. Initially, the consensus was that they were dimeric, based on the application of a large number of different techniques. However, the structures of the mitochondrial ADP/ATP carrier, a member of the family, clearly demonstrated that its structural fold is monomeric, lacking a conserved dimerisation interface. A re-evaluation of previously published data, with the advantage of hindsight, concluded that technical errors were at the basis of the earlier dimer claims. Here, we revisit this topic, as new claims for the existence of dimers of the bovine ADP/ATP carrier have emerged using native mass spectrometry of mitochondrial membrane vesicles. However, the measured mass does not agree with previously published values, and a large number of post-translational modifications are proposed to account for the difference. Contrarily, these modifications are not observed in electron density maps of the bovine carrier. If they were present, they would interfere with the structure and function of the carrier, including inhibitor and substrate binding. Furthermore, the reported mass does not account for three tightly bound cardiolipin molecules, which are consistently observed in other studies and are important stabilising factors for the transport mechanism. The monomeric carrier has all of the required properties for a functional transporter and undergoes large conformational changes that are incompatible with a stable dimerisation interface. Thus, our view that the native mitochondrial ADP/ATP carrier exists and functions as a monomer remains unaltered.

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Membrane Protein Production in Lactococcus lactis for Functional Studies

Cited by 2 publications

References 48 publications

Ectopic Neo-Formed Intracellular Membranes in Escherichia coli: A Response to Membrane Protein-Induced Stress Involving Membrane Curvature and Domains

Ectopic Neo-Formed Intracellular Membranes in Escherichia coli: A Response to Membrane Protein-Induced Stress Involving Membrane Curvature and Domains

The mitochondrial ADP/ATP carrier exists and functions as a monomer

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