A Solid-State NMR Index of Helical Membrane Protein Structure and Topology

Marassi, Francesca M.; Opella, Stanley J.

doi:10.1006/jmre.2000.2035

Cited by 338 publications

(406 citation statements)

References 29 publications

(27 reference statements)

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“…Images of proteins and peptides have been used to address various biological questions related to membrane-peptide and peptide-peptide interactions that determine the function of membrane-associated proteins (11,40,65). Our PISEMA results presented in this paper are consistent with the interpretations and explanations provided by the previous studies on different transmembrane helical systems (13,18,40,60,61,65). Interestingly, the tilt angles measured from solid-state NMR experiments are also in good agreement with MD simulations.…”

Section: Discussionsupporting

confidence: 89%

“…The simulations on the hydrophobic sequence (the TM peptide) alone suggest that the tilt angle is 19.8° and 24.6° in POPC and DMPC bilayers respectively. Solid-state NMR experiments have been successfully used to understand the topology of several membrane-associated peptides and proteins (13,(18)(19)(20)(21)40,60,61). Particularly, high resolution rendered by 2D PISEMA experiments has yielded PISA wheel patterns of membrane-bound helices (13,18,40,60,61).…”

Section: Discussionmentioning

confidence: 99%

“…Solid-state NMR experiments have been successfully used to understand the topology of several membrane-associated peptides and proteins (13,(18)(19)(20)(21)40,60,61). Particularly, high resolution rendered by 2D PISEMA experiments has yielded PISA wheel patterns of membrane-bound helices (13,18,40,60,61). Images of proteins and peptides have been used to address various biological questions related to membrane-peptide and peptide-peptide interactions that determine the function of membrane-associated proteins (11,40,65).…”

Section: Discussionmentioning

confidence: 99%

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Structure, Topology, and Tilt of Cell-Signaling Peptides Containing Nuclear Localization Sequences in Membrane Bilayers Determined by Solid-State NMR and Molecular Dynamics Simulation Studies

et al. 2007

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Cell-signaling peptides have been extensively used to transport functional molecules across the plasma membrane into living cells. These peptides consist of a hydrophobic sequence and a cationic nuclear localization sequence (NLS). It has been assumed that the hydrophobic region penetrates through the hydrophobic lipid bilayer and delivers the NLS inside the cell. To better understand the transport mechanism of these peptides, in this study, we investigated the structure, orientation, tilt of the peptide relative to the bilayer normal, and the membraneinteraction of two cell-signaling peptides, SA and SKP. Results from CD and solid-state NMR experiments combined with molecular dynamics simulations suggest that the hydrophobic region is helical and has a transmembrane orientation with the helical axis tilted away from the bilayer normal. The influence of the hydrophobic mismatch, between the hydrophobic length of the peptide and the hydrophobic thickness of the bilayer, on the tilt angle of the peptides was investigated using thicker POPC and thinner DMPC bilayers. NMR experiments showed that the hydrophobic domain of each peptide has a tilt angle of 15±3° in POPC, while in DMPC 25±3° and 30±3° tilts were observed for SA and SKP peptides respectively. These results are in good agreement with molecular dynamics simulations, which predicts a tilt angle of 13.3° (SA in POPC), 16.4° (SKP in POPC), 22.3° (SA in DMPC) and 31.7°( SKP in POPC). These results and simulations on the hydrophobic fragment of SA or SKP suggest that the tilt of helices increases with a decrease in the bilayer thickness without changing the phase, order, and structure of the lipid bilayers. KeywordsCell-permeable peptides; NLS; PISA wheel; tilt; hydrophobic mismatch; transmembrane helix Noninvasive delivery of peptides, proteins, oligonucleotides and other functional cargo molecules into living cells is highly dependent on their efficient transport across the plasma membrane barrier (1-7). Hydrophobic peptides have been successfully used to transport nuclear localization sequences (NLS) in order to probe intracellular signaling, which involved

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Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Structure, Topology, and Tilt of Cell-Signaling Peptides Containing Nuclear Localization Sequences in Membrane Bilayers Determined by Solid-State NMR and Molecular Dynamics Simulation Studies

et al. 2007

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“…1(d)]. 39 Using this approach, and by identifying the so-called PISA wheel patterns in 2D 15 N NMR spectra, 40,41 we have determined the orientation of the voltage-sensing S4 helix of a potassium chancel in lipid bicelles. 42 Although this oriented-sample approach is the mainstay of orientation determination by solid-state NMR, because of the dependence of the order parameters of uniaxially diffusing molecules on molecular orientation, one can now also determine protein orientation using unoriented liposomes.…”

Section: Solid-state Nmr Techniques For Studying Protein-membrane Intmentioning

confidence: 99%

Structure and dynamics of cationic membrane peptides and proteins: Insights from solid‐state NMR

Hong¹,

Su²

2011

Protein Science

130

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Many membrane peptides and protein domains contain functionally important cationic Arg and Lys residues, whose insertion into the hydrophobic interior of the lipid bilayer encounters significant energy barriers. To understand how these cationic molecules overcome the free energy barrier to insert into the lipid membrane, we have used solid-state NMR spectroscopy to determine the membrane-bound topology of these peptides. A versatile array of solid-state NMR experiments now readily yields the conformation, dynamics, orientation, depth of insertion, and site-specific protein-lipid interactions of these molecules. We summarize key findings of several Arg-rich membrane peptides, including b-sheet antimicrobial peptides, unstructured cell-penetrating peptides, and the voltage-sensing helix of voltage-gated potassium channels. Our results indicate the central role of guanidinium-phosphate and guanidinium-water interactions in dictating the structural topology of these cationic molecules in the lipid membrane, which in turn account for the mechanisms of this functionally diverse class of membrane peptides.

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“…2d) has several identifiable peaks at frequencies throughout the range of the 15 N amide chemical shift, and gives the first view of the structural organization of this protein in a membrane. Resonances around 200 ppm are from backbone amide sites in the single transmembrane helix that have their NH bonds nearly perpendicular to the plane of the membrane, and the rather narrow dispersion of 15 N resonances centered around 200 ppm suggests that the CHIF transmembrane helix crosses the lipid bilayer membrane with only a modest tilt angle [25,26]. Resonances around 80 ppm are from sites in the N-and C-terminal helices, with NH bonds nearly parallel to the membrane surface, and the peak near 35 ppm results from the amino groups of the lysine sidechains and the N-terminus.…”

Section: Membrane Proteins In Hydration-optimized Lipid Bilayer Samplesmentioning

confidence: 99%

Hydration-optimized oriented phospholipid bilayer samples for solid-state NMR structural studies of membrane proteins

Marassi

Crowell

2003

Journal of Magnetic Resonance

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The preparation of oriented, hydration-optimized lipid bilayer samples, for NMR structure determination of membrane proteins, is described. The samples consist of planar phospholipid bilayers, containing membrane proteins, that are oriented on single pairs of glass slides, and are placed in the coil of the NMR probe with the bilayer plane perpendicular to the direction of the magnetic field. Lipid bilayers provide a medium that closely resembles the biological membrane, and sample orientation both preserves the intrinsic membrane-defined directional quality of membrane proteins, and provides the mechanism for resonance line narrowing. The hydrationoptimized samples overcome some of the difficulties associated with multi-dimensional, highresolution, solid-state NMR spectroscopy of membrane proteins. These samples have greater stability over the course of multidimensional NMR experiments, they have lower sample conductance for greater rf power efficiency, and enable greater rf coil filling factors to be obtained for improved experimental sensitivity. Sample preparation is illustrated for the membrane protein CHIF (channel inducing factor), a member of the FXYD family of ion transport regulators.

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A Solid-State NMR Index of Helical Membrane Protein Structure and Topology

Cited by 338 publications

References 29 publications

Structure, Topology, and Tilt of Cell-Signaling Peptides Containing Nuclear Localization Sequences in Membrane Bilayers Determined by Solid-State NMR and Molecular Dynamics Simulation Studies

Structure, Topology, and Tilt of Cell-Signaling Peptides Containing Nuclear Localization Sequences in Membrane Bilayers Determined by Solid-State NMR and Molecular Dynamics Simulation Studies

Structure and dynamics of cationic membrane peptides and proteins: Insights from solid‐state NMR

Hydration-optimized oriented phospholipid bilayer samples for solid-state NMR structural studies of membrane proteins

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