Salt-Dependent Interactions between the C-Terminal Domain of Osmoregulatory Transporter ProP of <i>Escherichia coli</i> and the Lipid Membrane

Öztürk, Tuğba N.; Culham, Doreen E.; Tempelhagen, Laura; Wood, Janet M.; Lamoureux, Guillaume

doi:10.1021/acs.jpcb.0c03935

Cited by 5 publications

(14 citation statements)

References 52 publications

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“…Within the first 10 ns of simulation, the CTDs in both models began to interact with the membrane surface and these interactions were preserved throughout 10 μs-long simulations. This observation is consistent with previous simulations in which stable interactions between the membrane and the CTD were observed in structures modeled both with and without TM XII …”

Section: Resultssupporting

confidence: 93%

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Structural Determinants and Functional Significance of Dimerization for Osmosensing Transporter ProP in Escherichia coli

et al. 2022

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Osmosensing transporter ProP forestalls cellular dehydration by detecting environments with high osmotic pressure and mediating the accumulation of organic osmolytes by bacterial cells. It is composed of 12 transmembrane helices with cytoplasmic N-and C-termini. In Escherichia coli, dimers form when the C-terminal domains of ProP molecules form homodimeric, antiparallel, α-helical coiled coils. No dominant negative effect was detected when inactive and active ProP molecules formed heterodimers in vivo. Purification of ProP in detergent dodecylmaltoside yielded monomers, which were functional after reconstitution in proteoliposomes. With other evidence, this suggests that ProP monomers function independently whether in the monomeric or dimeric state. Amino acid replacements that disrupted or reversed the coiled coil did not prevent in vivo dimerization of ProP detected with a bacterial twohybrid system. Maleimide labeling detected no osmolality-dependent variation in the reactivities of cysteine residues introduced to transmembrane helix (TM) XII. In contrast, coarse-grained molecular dynamic simulations detected deformation of the lipid around TMs III and VI, on the lipid-exposed protein surface opposite to TM XII. This suggests that the dimer interface of ProP includes the surfaces of TMs III and VI, not of TM XII as previously suggested by crosslinking data. Homology modeling suggested that coiledcoil formation and dimerization via such an interface are not mutually exclusive. In previous work, alterations to the C-terminal coiled coil blocked co-localization of ProP with phospholipid cardiolipin at E. coli cell poles. Thus, dimerization may contribute to ProP targeting, adjust its lipid environment, and hence indirectly modify its osmotic stress response.

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Section: Resultssupporting

confidence: 93%

“…Second, hydrophobic mismatch and membrane thinning may occur when membrane-integral ProPEc surfaces constituting a dimer interface are exposed to phospholipid. Third, as discussed above, the osmotic activation of ProPEc is believed to involve interactions of the CTD α-helix with the membrane surface. ,, …”

Section: Resultsmentioning

confidence: 99%

Structural Determinants and Functional Significance of Dimerization for Osmosensing Transporter ProP in Escherichia coli

et al. 2022

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“…Osmosensing transporters like ProP and ProVWX activate to mediate osmolyte uptake as other transporters inactivate at high osmotic pressure [40]. It showed that E. coli protein ProP is both an osmosensor and an osmoprotectant transporter [41]. Therefore, ProP also plays an important role in the structure of membrane wall in E. coli.…”

Section: Discussionmentioning

confidence: 99%

Improving l-threonine production in Escherichia coli by elimination of transporters ProP and ProVWX

et al. 2021

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Background Betaine, an osmoprotective compatible solute, has been used to improve l-threonine production in engineered Escherichia colil-threonine producer. Betaine supplementation upregulates the expression of zwf encoding glucose-6-phosphate dehydrogenase, leading to the increase of NADPH, which is beneficial for l-threonine production. In E. coli, betaine can be taken through ProP encoded by proP or ProVWX encoded by proVWX. ProP is a H+-osmolyte symporter, whereas ProVWX is an ABC transporter. ProP and ProVWX mediate osmotic stress protection by transporting zwitterionic osmolytes, including glycine betaine. Betaine can also be synthesized in E. coli by enzymes encoded by betABIT. However, the influence of ProP, ProVWX and betABIT on l-threonine production in E. coli has not been investigated. Results In this study, the influence of ProP, ProVWX and betABIT on l-threonine production in E. coli has been investigated. Addition of betaine slightly improved the growth of the l-threonine producing E. coli strain TWF001 as well as the l-threonine production. Deletion of betABIT retarded the growth of TWF001 and slightly decreased the l-threonine production. However, deletion of proP or/and proVWX significantly increased the l-threonine production. When proP was deleted, the l-threonine production increased 33.3%; when proVWX was deleted, the l-threonine production increased 40.0%. When both proP and proVWX were deleted, the resulting strain TSW003 produced 23.5 g/l l-threonine after 36 h flask cultivation. The genes betABIT, proC, fadR, crr and ptsG were individually deleted from TSW003, and it was found that further absence of either crr (TWS008) or ptsG (TWS009) improved l-threonine production. TSW008 produced 24.9 g/l l-threonine after 36 h flask cultivation with a yield of 0.62 g/g glucose and a productivity of 0.69 g/l/h. TSW009 produced 26 g/l l-threonine after 48 h flask cultivation with a yield of 0.65 g/g glucose and a productivity of 0.54 g/l/h, which is 116% increase compared to the control TWF001. Conclusions In this study, l-threonine-producing E. coli strains TSW008 and TSW009 with high l-threonine productivity were developed by regulating the intracellular osmotic pressure. This strategy could be used to improve the production of other products in microorganisms.

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“…For example, transcription of the gene encoding the PutP ortholog OpuE in Bacillus subtilus is upregulated by means of sigma A-and sigma B-dependent stress-responsive promoters [40,41]. However, contrary to other proline and betaine transporters such as ProP (major facilitator superfamily [42]) and BetP (betaine/carnitine/choline transporter family [43], the activity of PutP and its orthologs is not stimulated at the protein level [44]. Furthermore, intracellular accumulation of L-proline can protect mammals, plants, fungi, yeast, and bacteria from damage by reactive oxygen species (for example, •OH, 1 O2) [45].…”

Section: Putp-mediated Nutrient Supply and Osmoadaptationmentioning

confidence: 99%

Prokaryotic Solute/Sodium Symporters: Versatile Functions and Mechanisms of a Transporter Family

Henríquez

Wirtz

et al. 2021

IJMS

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The solute/sodium symporter family (SSS family; TC 2.A.21; SLC5) consists of integral membrane proteins that use an existing sodium gradient to drive the uphill transport of various solutes, such as sugars, amino acids, vitamins, or ions across the membrane. This large family has representatives in all three kingdoms of life. The human sodium/iodide symporter (NIS) and the sodium/glucose transporter (SGLT1) are involved in diseases such as iodide transport defect or glucose-galactose malabsorption. Moreover, the bacterial sodium/proline symporter PutP and the sodium/sialic acid symporter SiaT play important roles in bacteria–host interactions. This review focuses on the physiological significance and structural and functional features of prokaryotic members of the SSS family. Special emphasis will be given to the roles and properties of proteins containing an SSS family domain fused to domains typically found in bacterial sensor kinases.

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Salt-Dependent Interactions between the C-Terminal Domain of Osmoregulatory Transporter ProP of Escherichia coli and the Lipid Membrane

Cited by 5 publications

References 52 publications

Structural Determinants and Functional Significance of Dimerization for Osmosensing Transporter ProP in Escherichia coli

Structural Determinants and Functional Significance of Dimerization for Osmosensing Transporter ProP in Escherichia coli

Improving l-threonine production in Escherichia coli by elimination of transporters ProP and ProVWX

Prokaryotic Solute/Sodium Symporters: Versatile Functions and Mechanisms of a Transporter Family

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