Electrostatics and Intrinsic Disorder Drive Translocon Binding of the SRP Receptor FtsY

Lakomek, Nils‐Alexander; Draycheva, Albena; Bornemann, Thomas; Wintermeyer, Wolfgang

doi:10.1002/anie.201602905

Cited by 15 publications

(17 citation statements)

References 28 publications

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“…Our previous results also showed that FtsY binds to SecYEG only when it is surrounded by phospholipids, as complex formation is not observed with SecYEG solubilized by adding detergent (Kuhn et al, 2015). FtsY binding to empty lipid nanodiscs is much weaker (K d 5 1.2 mM) (Kuhn et al, 2015), indicating that the affinity of the complex of FtsY with SecYEG embedded in a phospholipid bilayer is governed by the electrostatic interaction of FtsY with SecY (Lakomek et al, 2016), although there may be a contribution of anionic phospholipids, which are accumulated around the translocon (Prabudiansyah et al, 2015). An enrichment of anionic phospholipids we observe for nanodiscs containing SecYEG, as well (see "Experimental Procedures" section).…”

Section: Interaction Of Ftsy Domains With the Secyeg Transloconmentioning

confidence: 71%

“…The spectral resonances can be assigned to individual amino acids in FtsY-A207, as described elsewhere (Lakomek et al, 2016). Briefly, about 180 out of 190 expected resonances (207 amino acids minus 16 invisible proline residues and the invisible N-terminal amino acid) are visible in two-dimensional 15 N 1 H-TROSY-HSQC spectra of FtsY-A207; the remaining resonances either are hidden by spectral overlap or broadened below the detection threshold as the result of dynamic processes on the micro-to millisecond time scale.…”

Section: Nmr Spectroscopy Defines Interaction Site In the Mtsmentioning

confidence: 99%

“…The highly acidic A domain tethers FtsY to the membrane (de Leeuw et al, 1997;Zelazny et al, 1997;Weiche et al, 2008;Braig et al, 2009). The A domain is intrinsically disordered (Stjepanovic et al, 2011;Lakomek et al, 2016) and highly divergent in size among various prokaryotes (Bibi et al, 2001;Haddad et al, 2005). It preferentially binds to anionic phospholipids via two conserved membrane-binding sequences.…”

Section: Introductionmentioning

confidence: 99%

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The bacterial SRP receptor, FtsY, is activated on binding to the translocon

Draycheva

Bornemann

Ryazanov

et al. 2016

Molecular Microbiology

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SummaryProteins are inserted into the bacterial plasma membrane cotranslationally after translating ribosomes are targeted to the translocon in the membrane via the signal recognition particle (SRP) pathway. The targeting pathway involves an interaction between SRP and the SRP receptor, FtsY. Here we focus on the role of FtsY and its interaction with the translocon in controlling targeting. We show that in unbound FtsY the NG and A domains interact with one another. The interaction involves the membrane-targeting region at the junction between A and N domain. The closed form of FtsY is impaired in binding to SRP. Upon binding to the phospholipid-embedded translocon the domains of FtsY move apart. This enhances the docking of the FtsY NG domain to the homologous NG domain of the SRP protein Ffh. Thus, FtsY binding to the translocon has a central role in orchestrating the formation of a quaternary transfer complex in which the nascent peptide is transferred to the translocon. We propose that FtsY activation at the translocon ensures that ribosome-SRP complexes are directed to available translocons. This way sequestering SRP in futile complexes with unbound FtsY can be avoided and efficient targeting to the translocon achieved.

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Section: Interaction Of Ftsy Domains With the Secyeg Transloconmentioning

confidence: 71%

Section: Nmr Spectroscopy Defines Interaction Site In the Mtsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The bacterial SRP receptor, FtsY, is activated on binding to the translocon

Draycheva

Bornemann

Ryazanov

et al. 2016

Molecular Microbiology

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“…According to this model, the translocon is initially recruited to the targeting complex via hydrophobic and electrostatic interactions with the intrinsically disordered A-domain of SR232427. However, in spite of its central importance in the SRP cycle, such a quaternary complex has never been visualized and it is currently not understood how the final events of this process are spatially and temporally orchestrated.…”

mentioning

confidence: 99%

Structure of the quaternary complex between SRP, SR, and translocon bound to the translating ribosome

Jomaa

Boehringer

et al. 2017

Nat Commun

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During co-translational protein targeting, the signal recognition particle (SRP) binds to the translating ribosome displaying the signal sequence to deliver it to the SRP receptor (SR) on the membrane, where the signal peptide is transferred to the translocon. Using electron cryo-microscopy, we have determined the structure of a quaternary complex of the translating Escherichia coli ribosome, the SRP–SR in the ‘activated' state and the translocon. Our structure, supported by biochemical experiments, reveals that the SRP RNA adopts a kinked and untwisted conformation to allow repositioning of the ‘activated' SRP–SR complex on the ribosome. In addition, we observe the translocon positioned through interactions with the SR in the vicinity of the ribosome exit tunnel where the signal sequence is extending beyond its hydrophobic binding groove of the SRP M domain towards the translocon. Our study provides new insights into the mechanism of signal sequence transfer from the SRP to the translocon.

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“…[1] They play an important role in the binding of proteins to small ligands, [2] proteins, [3] DNA, [4] and other molecules. [5] It has also been shown that protein thermostability can be improved through optimizing the surface charge-charge interactions.…”

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confidence: 99%

Entropy Drives the Formation of Salt Bridges in the Protein GB3

Zhang

Wang

et al. 2017

Angewandte Chemie

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Salt bridges are very common in proteins.But what drives the formation of protein salt bridges is not clear.Inthis work, we determined the strength of four salt bridges in the protein GB3 by measuring the DpK a values of the basic residues that constitute the salt bridges with ah ighly accurate NMR titration method at different temperatures.T he results show that the DpK a values increase with temperature,t hus indicating that the salt bridges are stronger at higher temperatures.Fitting of DpK a values to the vantHoff equation yields positive DHand DSvalues,thus indicating that entropydrives salt-bridge formation. Molecular dynamics simulations show that the protein and solvent make opposite contributions to DH and DS. Specifically,t he enthalpic gain contributed from the protein is more than offset by the enthalpic loss contributed from the solvent, whereas the entropic gain originates from the desolvation effect.

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Electrostatics and Intrinsic Disorder Drive Translocon Binding of the SRP Receptor FtsY

Cited by 15 publications

References 28 publications

The bacterial SRP receptor, FtsY, is activated on binding to the translocon

The bacterial SRP receptor, FtsY, is activated on binding to the translocon

Structure of the quaternary complex between SRP, SR, and translocon bound to the translating ribosome

Entropy Drives the Formation of Salt Bridges in the Protein GB3

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