The Native Conformation of the Human VDAC1 N Terminus

Schneider, Robert; Etzkorn, Manuel; Giller, Karin; Daebel, Venita; Eisfeld, Jörg; Zweckstetter, Markus; Griesinger, Christian; Becker, Stefan; Lange, Adam

doi:10.1002/anie.200906241

Cited by 63 publications

(120 citation statements)

References 23 publications

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“…In total, five residues are assigned in the second half of the N-terminal helix (residues [12][13][14][15][16][17][18][19][20], demonstrating that this region is not broadened beyond detection by intermediate chemical exchange. This result is in line with the observation that peaks corresponding to residues A2-V17 give rise to distinct narrow crosspeaks in solid state NMR spectra of hVDAC1 reconstituted in liposomes, indicating a rather rigid conformation (24).…”

Section: μS-ms Dynamics Are Significantly Increased For the N-terminasupporting

confidence: 90%

“…S4). Such a stabilizing role of the helix has also been suggested on the basis of solid-state NMR results on an N-terminally truncated version of hVDAC1 (24). In contrast, high flexibility was found from β2 to the top end of β7 (Fig.…”

Section: μS-ms Dynamics Are Significantly Increased For the N-terminasupporting

confidence: 52%

“…NMR spectra were recorded on 2 Hð75%Þ-15 N or 2 Hð75%Þ-13 C-15 N labeled samples containing 0.5-0.8 mM hVDAC1, 25 mM BisTris pH 6.8, approximately 250 mM LDAO (Fluka), and 5-10% D 2 O. Improvement of backbone assignment was achieved by TROSY-HNCA experiments (56), 1 H, 15 N-TROSY experiments (57) at varying temperatures and C α shifts from solid-state NMR experiments (24). Backbone dynamics were determined from steady state heteronuclear f 1 Hg, 15 N-NOEs (25) and chemical exchange rates (R ex ) (26).…”

Section: Methodsmentioning

confidence: 99%

“…With improved sample quality and additional NMR experiments we now increased the assignment to 84/97% (overall/barrel). The assignment of three residues in the N-terminal helix (K12, D16, and V17) benefited from the chemical shift information obtained from solidstate NMR (24). In total, five residues are assigned in the second half of the N-terminal helix (residues [12][13][14][15][16][17][18][19][20], demonstrating that this region is not broadened beyond detection by intermediate chemical exchange.…”

Section: μS-ms Dynamics Are Significantly Increased For the N-terminamentioning

confidence: 99%

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Functional dynamics in the voltage-dependent anion channel

Villinger

Briones

Giller

et al. 2010

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

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105

129

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The voltage-dependent anion channel (VDAC), located in the outer mitochondrial membrane, acts as a gatekeeper for the entry and exit of mitochondrial metabolites. Here we reveal functional dynamics of isoform one of VDAC (VDAC1) by a combination of solution NMR spectroscopy, Gaussian network model analysis, and molecular dynamics simulation. Micro-to millisecond dynamics are significantly increased for the N-terminal six β-strands of VDAC1 in micellar solution, in agreement with increased B-factors observed in the same region in the bicellar crystal structure of VDAC1. Molecular dynamics simulations reveal that a charge on the membrane-facing glutamic acid 73 (E73) accounts for the elevation of N-terminal protein dynamics as well as a thinning of the nearby membrane. Mutation or chemical modification of E73 strongly reduces the micro-to millisecond dynamics in solution. Because E73 is necessary for hexokinase-I-induced VDAC channel closure and inhibition of apoptosis, our results imply that microto millisecond dynamics in the N-terminal part of the barrel are essential for VDAC interaction and gating.membrane protein | molecular dynamics | NMR spectroscopy | protein-lipid interactions | structure D ynamics of proteins provide the essential link between structure and function. Membrane protein dynamics are gaining more attention due to an increasing number of high-resolution structures. A versatile experimental method for the study of membrane protein dynamics is NMR spectroscopy, providing atomic resolution information from nanoseconds to seconds (for an overview see ref. 1). In addition, when a three-dimensional structure is available, insight into fast dynamics of proteins, which occur on the nanosecond time scale, might be gained from molecular dynamics (MD) simulation and elastic network models (2, 3).NMR studies have revealed large conformational changes essential for the physiological function of α-helical membrane proteins, such as the interaction of phospholamban with effectors modulating heart muscle contractility (4, 5) and antimicrobial peptides adopting different conformations in membranes or solution (6, 7). Less well characterized are motions of outer membrane β-barrels. A pioneering solution NMR study revealed μs-ms interconversion of the catalytic loop of PagP between an excited and a nonexcited state, enabling both ligand binding and enzymatic catalysis (8). Nonenzymatic β-barrel channels exhibit more general dynamic features, such as loop flexibility and slow conformational exchange towards the extracellular barrel edges, altogether indicating relevance for substrate conductance and gating (9-11).The voltage-dependent anion channel (VDAC) is one of the few β-barrel membrane proteins, the structure of which has been determined to date (12-14). Serving as the major pass-through for metabolites between mitochondria and the cytosol (15), the highly abundant channel is regarded as a key regulator of cellular energy metabolism (16). Metabolite flow is regulated by a voltage-dependent conformational c...

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Section: μS-ms Dynamics Are Significantly Increased For the N-terminasupporting

confidence: 90%

Section: μS-ms Dynamics Are Significantly Increased For the N-terminasupporting

confidence: 52%

Section: Methodsmentioning

confidence: 99%

Section: μS-ms Dynamics Are Significantly Increased For the N-terminamentioning

confidence: 99%

See 2 more Smart Citations

Functional dynamics in the voltage-dependent anion channel

Villinger

Briones

Giller

et al. 2010

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

Self Cite

105

129

View full text Add to dashboard Cite

show abstract

“…These data were supported recently by a solid-state NMR analysis of hVDAC1 in liposomes demonstrating that the general fold of hVDAC1 is also maintained in a more native-like lipid membrane environment. 12 Several models for the voltage-gating mechanism were based on rearrangements of these unique structural elements of the a-helix and the parallel b-sheet. [13][14][15][16] These theories involved, in particular, movements of the helix into the centre of the pore or realignments of the b-sheet structure to change the pore size or shape, 13,14 or the electrical properties of the lumen of VDAC.…”

Section: Introductionmentioning

confidence: 99%

Voltage-dependent structural changes of the membrane-bound anion channel hVDAC1 probed by SEIRA and electrochemical impedance spectroscopy

Kozuch

Weichbrodt

Millo

et al. 2014

Phys. Chem. Chem. Phys.

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The voltage-dependent anion channel (VDAC) is a transmembrane protein that regulates the transfer of metabolites between the cytosol and the mitochondrium. Opening and partial closing of the channel is known to be driven by the transmembrane potential via a mechanism that is not fully understood. In this work, we employed a spectroelectrochemical approach to probe the voltage-induced molecular structure changes of human VDAC1 (hVDAC1) embedded in a tethered bilayer lipid membrane on a nanostructured Au electrode. The model membrane consisted of a mixed self-assembled monolayer of 6-mercaptohexanol and (cholesterylpolyethylenoxy)thiol, followed by the deposition of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine vesicles including hVDAC1. The stepwise assembly of the model membrane and the incorporation of hVDAC1 were monitored by surface enhanced infrared absorption and electrochemical impedance spectroscopy. Difference spectra allowed for identifying the spectral changes which may be associated with the transition from the open to the ''closed'' states by shifting the potential above or below the transmembrane potential determined to be ca. 0.0 V vs. the open circuit potential. These spectral changes were interpreted on the basis of the orientation-and distancedependent IR enhancement and indicate alterations of the inclination angle of the b-strands as crucial molecular events, reflecting an expansion or contraction of the b-barrel pore. These protein structural changes that do not confirm nor exclude the reorientation of the a-helix are either directly induced by the electric field or a consequence of a potential-dependent repulsion or attraction of the bilayer.

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Lipid Dynamics and Protein-Lipid Interactions in Integral Membrane Proteins: Insights from Solid-State NMR

Wel

2014

eMagRes

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The Native Conformation of the Human VDAC1 N Terminus

Cited by 63 publications

References 23 publications

Functional dynamics in the voltage-dependent anion channel

Functional dynamics in the voltage-dependent anion channel

Voltage-dependent structural changes of the membrane-bound anion channel hVDAC1 probed by SEIRA and electrochemical impedance spectroscopy

Lipid Dynamics and Protein-Lipid Interactions in Integral Membrane Proteins: Insights from Solid-State NMR

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