Solution Structure of the C-terminal Antiparallel Coiled-coil Domain from Escherichia coli Osmosensor ProP

Zoetewey, David; Tripet, Brian; Kutateladze, Tatiana G.; Overduin, Michael; Wood, Janet M.; Hodges, Robert S.

doi:10.1016/j.jmb.2003.10.020

Cited by 36 publications

(74 citation statements)

References 65 publications

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“…Interestingly, the solution structure of the C-terminal region of the osmosensor ProP revealed a coiled-coil domain [31], which is thought to be important for osmotic activation of ProP [32].…”

Section: Possible Role For N-alpha Domain In Osmosensingmentioning

confidence: 99%

Structural characterization of Escherichia coli sensor histidine kinase EnvZ: the periplasmic C-terminal core domain is critical for homodimerization

Khorchid

Inouye

Ikura

2004

Biochemical Journal

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Escherichia coli EnvZ is a membrane sensor histidine kinase that plays a pivotal role in cell adaptation to changes in extracellular osmolarity. Although the cytoplasmic histidine kinase domain of EnvZ has been extensively studied, both biochemically and structurally, little is known about the structure of its periplasmic domain, which has been implicated in the mechanism underlying its osmosensing function. In the present study, we report the biochemical and biophysical characterization of the periplasmic region of EnvZ (Ala 38 -Arg 162 ). This region was found to form a dimer in solution, and to consist of two well-defined domains: an N-terminal α-helical domain and a C-terminal core domain (Glu 83 -Arg 162 ) containing both α-helical and β-sheet secondary structures. Our pull-down assays and analytical ultracentrifugation analysis revealed that dimerization of the periplasmic region is highly sensitive to the presence of CHAPS, but relatively insensitive to salt concentration, thus suggesting the significance of hydrophobic interactions between the homodimeric subunits. Periplasmic homodimerization is mediated predominantly by the C-terminal core domain, while a regulatory function may be attributed mainly to the N-terminal α-helical domain, whose mutations have been shown previously to produce a high-osmolarity phenotype.

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Section: Possible Role For N-alpha Domain In Osmosensingmentioning

confidence: 99%

Structural characterization of Escherichia coli sensor histidine kinase EnvZ: the periplasmic C-terminal core domain is critical for homodimerization

Khorchid

Inouye

Ikura

2004

Biochemical Journal

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“…In this respect, ProP of E. coli may seem to behave more similarly to BetT than to BetP. For ProP, it is reported that an amino acid substitution (Arg-488-Ile) which hampers the formation of an antiparallel coiled-coil structure in its C-terminal cytoplasmic domain causes the transporter to be less sensitive to and only transiently activated by osmotic stress in vivo (Culham et al, 2000;Zoetewey et al, 2003). It is also noteworthy that the E. coli CaiT transporter of the BCCT family has a deduced C-terminal extension of only ten residues.…”

Section: Discussionmentioning

confidence: 99%

“…ProU is an ATP-binding cassette (ABC) transporter (May et al, 1986), whereas ProP is a secondary transporter driven by the proton-motive force. ProP belongs to the major facilitator superfamily and possesses a large cytoplasmic Cterminal domain (Culham et al, 2000;Zoetewey et al, 2003). These transporters are osmotically activated at both the transcriptional and post-translational levels.…”

Section: Introductionmentioning

confidence: 99%

Membrane topology and mutational analysis of the osmotically activated BetT choline transporter of Escherichia coli

Tøndervik

Strøm

2007

Microbiology

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For osmoprotection, Escherichia coli can synthesize glycine betaine from externally supplied choline by the Bet system (betTIBA products). The major carrier of choline is the high-affinity, proton-driven, secondary transporter BetT, which belongs to the BCCT family of transporters. Fusion proteins consisting of N-terminal fragments of BetT linked to b-galactosidase (LacZ) or alkaline phosphatase (PhoA) were constructed. By analysis of 51 fusion proteins with 37 unique fusion-points, the predictions that BetT comprised 12 membrane-spanning regions and that its N-and C-terminal extensions of about 12 and 180 amino acid residues, respectively, were situated in the cytoplasm were confirmed. This is believed to represent the first experimental examination of the membrane topology of a BCCT family protein. Osmotic upshock experiments were performed with spectinomycin-treated E. coli cells that had expressed the wild-type or a mutant BetT protein during growth at low osmolality (160 mosmol kg "1 ). The choline transport activity of wild-typeBetT increased tenfold when the cells were stressed with 0.4 M NaCl (total osmolality 780 mosmol kg

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“…The solution structure of the 1R48 antiparallel dimer shows ARG24a and a pair of ASP residues (ASP14e and ASP11b) in the neighboring helix within range for hydrogen bond formation. 9 The interactions of ARG24a in our minimized average model are summarized in Figure 1(A). In this model, ARG24a adopts a rotameric state that allows it to interact only with ASP14e.…”

Section: Dimersmentioning

confidence: 99%

“…The 1R48 dimer has a relatively large RMSD because the minimized average NMR structure is bent toward the b/e surface [9] while our model was built with ideal coiled-coil parameters. Typically, the RMSD converged to the final value within 0.5 ns.…”

Section: Generation Of Coiled-coil Structuresmentioning

confidence: 99%

Computational analysis of residue contributions to coiled‐coil topology

Torre

Lazaridis

2011

Protein Science

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A variety of features are thought to contribute to the oligomeric and topological specificity of coiled coils. In previous work, we examined the determinants of oligomeric state. Here, we examine the energetic basis for the tendency of six coiled-coil peptides to align their a-helices in antiparallel orientation using molecular dynamics simulations with implicit solvation (EEF1.1). We also examine the effect of mutations known to disrupt the topology of these peptides. In agreement with experiment, ARG or LYS at a or d positions were found to stabilize the antiparallel configuration. The modeling suggests that this is not due to a-a 0 or d-d 0 repulsions but due to interactions with e 0 and g 0 residues. TRP at core positions also favors the antiparallel configuration. Residues that disfavor parallel dimers, such as ILE at d, are better tolerated in, and thus favor the antiparallel configuration. Salt bridge networks were found to be more stabilizing in the antiparallel configuration for geometric reasons: antiparallel helices point amino acid side chains in opposite directions. However, the structure with the largest number of salt bridges was not always the most stable, due to desolvation and configurational entropy contributions. In tetramers, the extent of stabilization of the antiparallel topology by core residues is influenced by the e 0 residue on a neighboring helix. Residues at b and c positions in some cases also contribute to stabilization of antiparallel tetramers. This work provides useful rules toward the goal of designing coiled coils with a well-defined and predictable three-dimensional structure.

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Solution Structure of the C-terminal Antiparallel Coiled-coil Domain from Escherichia coli Osmosensor ProP

Cited by 36 publications

References 65 publications

Structural characterization of Escherichia coli sensor histidine kinase EnvZ: the periplasmic C-terminal core domain is critical for homodimerization

Structural characterization of Escherichia coli sensor histidine kinase EnvZ: the periplasmic C-terminal core domain is critical for homodimerization

Membrane topology and mutational analysis of the osmotically activated BetT choline transporter of Escherichia coli

Computational analysis of residue contributions to coiled‐coil topology

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