Comparison of the dynamics of the membrane‐bound form of fd coat protein in micelles and in bilayers by solution and solid‐state nitrogen‐15 nuclear magnetic resonance spectroscopy

Bogusky, Michael J.; Leo, Gregory C.; Opella, Stanley J.

doi:10.1002/prot.340040205

Cited by 27 publications

(28 citation statements)

References 32 publications

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“…In contrast, solid-state NMR using oriented membrane protein samples can give molecular details regarding both backbone structure and topology. Because a number of membrane protein structures recently determined show similar folds in micelles, lipid bilayers, and crystals (34,(37)(38)(39)(40)(41), we propose to combine the restraints from these techniques into a unique protocol with the goal of obtaining the highresolution structure (backbone and side chains) and topology within the lipid bilayer. In the literature, there are several examples of backbone structure determination of membrane protein using solid-state NMR data alone (42,43) and a few examples reported for the combined use of solution and solid-state NMR information in a qualitative fashion (39, 44, 45).…”

Section: Discussionmentioning

confidence: 99%

“…In fact, the L-shaped membrane architecture of PLN is reminiscent of the structures and topologies of the FXYD proteins (a family of membrane associated proteins that serve as subunits to Na,K-ATPases) (40), the fd coat protein (responsible for viral assembly) (56), the VpU protein from HIV-1 (57), and the M2 channel from the inf luenza A virus (58). Amphipathic helix motifs are known to be functionally important in a number of capacities, ranging from stabilizing protein structures to sensing changes in the physical properties of the membrane (59,60).…”

Section: Discussionmentioning

confidence: 99%

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Structure and topology of monomeric phospholamban in lipid membranes determined by a hybrid solution and solid-state NMR approach

Traaseth

Shi²,

Verardi

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

158

194

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Phospholamban (PLN) is an essential regulator of cardiac muscle contractility. The homopentameric assembly of PLN is the reservoir for active monomers that, upon deoligomerization form 1:1 complexes with the sarco(endo)plasmic reticulum Ca 2؉ -ATPase (SERCA), thus modulating the rate of calcium uptake. In lipid bilayers and micelles, monomeric PLN exists in equilibrium between a bent (or resting) T state and a more dynamic (or active) R state. Here, we report the high-resolution structure and topology of the T state of a monomeric PLN mutant in lipid bilayers, using a hybrid of solution and solid-state NMR restraints together with molecular dynamics simulations in explicit lipid environments. Unlike the previous structural ensemble determined in micelles, this approach gives a complete picture of the PLN monomer structure in a lipid bilayer. This hybrid ensemble exemplifies the tilt, rotation, and depth of membrane insertion, revealing the interaction with the lipids for all protein domains. The N-terminal amphipathic helical domain Ia (residues 1-16) rests on the surface of the lipid membrane with the hydrophobic face of domain Ia embedded in the membrane bilayer interior. The helix comprised of domain Ib (residues 23-30) and transmembrane domain II (residues 31-52) traverses the bilayer with a tilt angle of Ϸ24°. The specific interactions between PLN and lipid membranes may represent an additional regulatory element of its inhibitory function. We propose this hybrid method for the simultaneous determination of structure and topology for membrane proteins with compact folds or proteins whose spatial arrangement is dictated by their specific interactions with lipid bilayers.hybrid method ͉ membrane proteins ͉ oriented solid-state NMR ͉ molecular modeling ͉ PISEMA S tructure and topology are central to membrane protein function (1). Recently determined high-resolution structures reveal the compact folds for several membrane proteins, such as electron and proton-conducting proteins involved in photosynthesis and respiration (http://blanco.biomol.uci.edu/ Membrane Proteins xtal.html). However, a significant population of membrane proteins does not possess a compact tertiary fold, but has its fold space defined through interactions of secondary structure elements (helices, turns, and loops) with the lipid membrane, i.e., the topology (1). This is the case for phospholamban (PLN), a mammalian protein that is essential in the regulation of cardiac muscle contractility (2), and that has recently become a major target for gene therapy to ameliorate cardiomyopathies (3, 4). PLN is located in the sarco(endo)plasmic reticulum (SR) of cardiac myocytes, inhibiting the SR Ca 2ϩ -ATPase (SERCA) by shifting its relative Ca 2ϩ affinity (5). In vitro and in vivo experiments have shown PLN to exist as a homopentamer that deoligomerizes into active monomers that bind SERCA in a 1:1 molar ratio (6). The monomeric form of PLN exists in equilibrium between a dynamically disordered R state and a more restricted T state (7,8) and has 3 ...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Structure and topology of monomeric phospholamban in lipid membranes determined by a hybrid solution and solid-state NMR approach

Traaseth

Shi²,

Verardi

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

158

194

View full text Add to dashboard Cite

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“…It is generally assumed that the insertion of the protein in a lipid bilayer is accompanied with a major structural rearrangement that splits the continuous ␣-helix in the phage particle into an amphipathic and a transmembrane helix perpendicular to each other (Almeida and Opella, 1997;Bogusky et al, 1988;Bogusky et al, 1987;Henry and Sykes, 1992;Henry et al, 1987;Leo et al, 1987;Marvin, 1998;McDonnell et al, 1993). However, several lines of evidence in recent studies propose that the change in the secondary structure on going from the phage to a matching membrane Fig.…”

Section: Structure Of Bacteriophage M13 Major Coat Protein Majormentioning

confidence: 96%

Anchoring mechanisms of membrane-associated M13 major coat protein

Stopar

Spruijt

Hemminga

2006

Chemistry and Physics of Lipids

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“…The membrane‐bound form of the major pVIII coat protein of the filamentous fd bacteriophage resides within the inner membrane of infected Escherichia coli before incorporation into virus particles that are extruded through the cell membrane. The structure of the membrane‐bound form of the protein has been extensively studied in micelle samples by solution NMR spectroscopy (Cross and Opella 1980; Henry and Sykes 1992; McDonnell et al 1993; Van de Ven et al 1993; Williams et al 1996; Almeida and Opella 1997; Papavoine et al 1998), as well as with solid‐state NMR experiments on bilayer samples (Leo et al 1987; Bogusky et al 1988; Marassi et al 1997). The results are in agreement in showing that the protein has two distinct α‐helical segments, a short amphipathic in‐plane (IP) helix that rests on the membrane surface, a longer hydrophobic transmembrane (TM) helix, and few mobile residues near the N and C termini.…”

mentioning

confidence: 99%

Simultaneous assignment and structure determination of a membrane protein from NMR orientational restraints

2003

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A solid-state NMR approach for simultaneous resonance assignment and three-dimensional structure determination of a membrane protein in lipid bilayers is described. The approach is based on the scattering, hence the descriptor "shotgun," of 15 N-labeled amino acids throughout the protein sequence (and the resulting NMR spectra). The samples are obtained by protein expression in bacteria grown on media in which one type of amino acid is labeled and the others are not. Shotgun NMR short-circuits the laborious and time-consuming process of obtaining complete sequential assignments prior to the calculation of a protein structure from the NMR data by taking advantage of the orientational information inherent to the spectra of aligned proteins. As a result, it is possible to simultaneously assign resonances and measure orientational restraints for structure determination. A total of five two-dimensional 1 H/ 15 N PISEMA (polarization inversion spin exchange at the magic angle) spectra, from one uniformly and four selectively 15 N-labeled samples, were sufficient to determine the structure of the membrane-bound form of the 50-residue major pVIII coat protein of fd filamentous bacteriophage. Pisa (polarity index slat angle) wheels are an essential element in the process, which starts with the simultaneous assignment of resonances and the assembly of isolated polypeptide segments, and culminates in the complete three-dimensional structure of the protein with atomic resolution. The principles are also applicable to weakly aligned proteins studied by solution NMR spectroscopy.[The structure we determined for the membrane-bound form of the Fd bacteriophage pVIII coat protein has been deposited in the Protein Data Bank as PDB file 1MZT.]

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Comparison of the dynamics of the membrane‐bound form of fd coat protein in micelles and in bilayers by solution and solid‐state nitrogen‐15 nuclear magnetic resonance spectroscopy

Cited by 27 publications

References 32 publications

Structure and topology of monomeric phospholamban in lipid membranes determined by a hybrid solution and solid-state NMR approach

Structure and topology of monomeric phospholamban in lipid membranes determined by a hybrid solution and solid-state NMR approach

Anchoring mechanisms of membrane-associated M13 major coat protein

Simultaneous assignment and structure determination of a membrane protein from NMR orientational restraints

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