Mechanistic studies of the biogenesis and folding of outer membrane proteins in vitro and in vivo: What have we learned to date?

McMorran, Lindsay M.; Brockwell, David J.; Radford, Sheena E.

doi:10.1016/j.abb.2014.02.011

Cited by 45 publications

(46 citation statements)

References 239 publications

(520 reference statements)

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“…For comparison, in the conventional model (as summarized in panel B, Fig. 7), translocation, transportation, and membrane of the nascent ␤-barrel OMPs were usually considered as largely segregated events (1,5,20,28), which would result in the undesirable exposure of hydrophobic nascent OMPs in the hydrophilic periplasmic environment. Another evident advantage of forming such a supercomplex is that it allows energy to be directly supplied, via the molecular motor SecA, from the cytoplasm to the ␤-barrel OMP biogenesis process (as illustrated in Fig.…”

Section: Discussionmentioning

confidence: 99%

A Supercomplex Spanning the Inner and Outer Membranes Mediates the Biogenesis of β-Barrel Outer Membrane Proteins in Bacteria

Wang

Feng

et al. 2016

Journal of Biological Chemistry

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␤-barrel outer membrane proteins (OMPs) are ubiquitously present in Gram-negative bacteria, mitochondria and chloroplasts, and function in a variety of biological processes. The mechanism by which the hydrophobic nascent ␤-barrel OMPs are transported through the hydrophilic periplasmic space in bacterial cells remains elusive. Here, mainly via unnatural amino acid-mediated in vivo photo-crosslinking studies, we revealed that the primary periplasmic chaperone SurA interacts with nascent ␤-barrel OMPs largely via its N-domain but with ␤-barrel assembly machine protein BamA mainly via its satellite P2 domain, and that the nascent ␤-barrel OMPs interact with SurA via their N-and C-terminal regions. Additionally, via dual in vivo photo-crosslinking, we demonstrated the formation of a ternary complex involving ␤-barrel OMP, SurA, and BamA in cells. More importantly, we found that a supercomplex spanning the inner and outer membranes and involving the BamA, BamB, SurA, PpiD, SecY, SecE, and SecA proteins appears to exist in living cells, as revealed by a combined analyses of sucrose-gradient ultra-centrifugation, Blue native PAGE and mass spectrometry. We propose that this supercomplex integrates the translocation, transportation, and membrane insertion events for ␤-barrel OMP biogenesis.

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Section: Discussionmentioning

confidence: 99%

A Supercomplex Spanning the Inner and Outer Membranes Mediates the Biogenesis of β-Barrel Outer Membrane Proteins in Bacteria

Wang

Feng

et al. 2016

Journal of Biological Chemistry

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“…It was shown by solid-state NMR that the soluble POTRA domains and the membrane-embedded part of BamA show significant differences in their respective mobilities [324] within precipitated samples, but that these POTRA-movements are significantly restricted when the protein is reconstituted in proteoliposomes [319]. Contributions by NMR on other translocases and insertases have so far remained sparse and the interested reader is referred to excellent recent reviews [23,59,325,326]. …”

Section: Bamamentioning

confidence: 99%

Chaperones and chaperone–substrate complexes: Dynamic playgrounds for NMR spectroscopists

Burmann¹,

Hiller²

2015

Progress in Nuclear Magnetic Resonance Spectroscopy

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“…Inner membrane and the inner leaflet of the outer membrane are mainly composed by phosphatidylethanolamine followed by phosphatidylglycerol and few cardiolipin and the outer leaflet of the outer membrane is highly enriched in Lipopolysaccharide (Bernaudat et al 2011;Midgett and Madden 2007;McMorran et al 2014;Fernández and Vega 2016) Pichia pastoris Methylotrophic yeast; GRAS organism Efficient protein secretion with low levels of endogenous proteins. Capable of performing many PTM; low cost of culture media; industry-scale fermentation Glycosylation pattern different from mammalian; intracellular recovery of large amount of cells may require specific equipment (French-press); high oxygen demand Improved glyco-engineered strains obtained using the GlycoSwitch® technology; wide range of genetic tools, plasmids, strains, and promoters available; the preference for the respiratory growth allow to be cultivated at high cell densities.…”

Section: Escherichia Colimentioning

confidence: 99%

Smoothing membrane protein structure determination by initial upstream stage improvements

Pedro

Queiroz

Passarinha

2019

Appl Microbiol Biotechnol

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Membrane proteins (MP) constitute 20-30% of all proteins encoded by the genome of various organisms and perform a wide range of essential biological functions. However, despite they represent the largest class of protein drug targets, a relatively small number high-resolution 3D structures have been obtained yet. Membrane protein biogenesis is more complex than that of the soluble proteins and its recombinant biosynthesis has been a major drawback, thus delaying their further structural characterization. Indeed, the major limitation in structure determination of MP is the low yield achieved in recombinant expression, usually coupled to low functionality, pinpointing the optimization target in recombinant MP research. Recently, the growing attention that have been dedicated to the upstream stage of MP bioprocesses allowed great advances, permitting the evolution of the number of MP solved structures. In this review, we analyse and discuss effective solutions and technical advances at the level of the upstream stage using prokaryotic and eukaryotic organisms foreseeing an increase in expression yields of correctly folded MP and that may facilitate the determination of their three-dimensional structure. A section on techniques used to protein quality control and further structure determination of MP is also included. Lastly, a critical assessment of major factors contributing for a good decision-making process related to the upstream stage of MP is presented.

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Mechanistic studies of the biogenesis and folding of outer membrane proteins in vitro and in vivo: What have we learned to date?

Cited by 45 publications

References 239 publications

A Supercomplex Spanning the Inner and Outer Membranes Mediates the Biogenesis of β-Barrel Outer Membrane Proteins in Bacteria

A Supercomplex Spanning the Inner and Outer Membranes Mediates the Biogenesis of β-Barrel Outer Membrane Proteins in Bacteria

Chaperones and chaperone–substrate complexes: Dynamic playgrounds for NMR spectroscopists

Smoothing membrane protein structure determination by initial upstream stage improvements

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