Oriented Attachment and Membrane Reconstitution of His-Tagged Cytochrome<i>c</i>Oxidase to a Gold Electrode:  In Situ Monitoring by Surface-Enhanced Infrared Absorption Spectroscopy

Ataka, Kenichi; Giess, Frank; Knoll, Wolfgang; Naumann, Renate; Haber‐Pohlmeier, Sabina; Richter, Björn; Heberle, Joachim

doi:10.1021/ja045951h

Cited by 271 publications

(354 citation statements)

References 45 publications

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“…The rough gold surface used for SEIRA spectroscopy was modified by nickel chelating nitrilo-triacetic acid (Ni-NTA) according to published procedures (12,14,34). Each modification step was followed in situ by SEIRAS.…”

Section: Methodsmentioning

confidence: 99%

“…Binding of SR II via the C-terminal His-tag to the Ni-NTA modified gold surface was carried out by incubating a 6 M solution of protein in 0.05% (wt/vol) n-dodecyl-␤-D-maltopyranoside (DDM) (pH 5.8, 20°C, 4 M NaCl, 50 mM phosphate buffer, in the case of SRII from H. s.) or a 4.0 M solution of protein in 0.02% DDM (pH 8.0, 20°C, 0.5M NaCl, 10 mM bis-Tris-propane buffer, in the case of SRII from N. p.) atop the Ni-NTA modified gold surface. Reconstitution of the membrane protein in the lipid bilayer by detergent removal (12) and the parallel addition of polar lipids from the purple membrane of H. s. (35) or phosphatidylcholine from egg yolk (Sigma) at a protein/lipid ratio of 1:20 stabilized the protein to achieve a good signal-to-noise ratio in the IR difference spectra by extensive signal averaging (vide infra). Saturation of protein binding to the modified surface occurred within Ϸ1.5 h. For light-induced IR difference spectroscopy, the protein was excited through a fiber optic by light from a 150 W tungsten lamp filtered by a narrow band filter (480 -505 nm) in the case of SRII from N. p. or by a light-emitting diode (LED; Luxeon Star) with the emission maximum at 497 nm (23 nm FWHM) and an intensity of 10 mW/cm 2 in the case of SRII from H. s.…”

Section: Methodsmentioning

confidence: 99%

“…The use of affinity tags yields orientated protein molecules along the solid surface. Importantly, surface-adhered membrane proteins are reconstituted directly at the surface into a lipid bilayer by detergent removal (12,13). The high surface sensitivity renders SEIRAS an exquisite method for studies of monolayers of surface-attached proteins.…”

mentioning

confidence: 99%

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Resolving voltage-dependent structural changes of a membrane photoreceptor by surface-enhanced IR difference spectroscopy

Jiang

Zaitseva

Schmidt

et al. 2008

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

129

161

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Membrane proteins are molecular machines that transport ions, solutes, or information across the cell membrane. Electrophysiological techniques have unraveled many functional aspects of ion channels but suffer from the lack of structural sensitivity. Here, we present spectroelectrochemical data on vibrational changes of membrane proteins derived from a single monolayer. ion transfer ͉ membrane potential ͉ proton translocation ͉ vibrational spectroscopy ͉ sensory rhodopsin

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Resolving voltage-dependent structural changes of a membrane photoreceptor by surface-enhanced IR difference spectroscopy

Jiang

Zaitseva

Schmidt

et al. 2008

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

129

161

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“…Recent advances in methodology and interpretation have produced insights into the catalytic mechanism in several biological systems (for examples, see refs. [1][2][3][4].…”

mentioning

confidence: 99%

Time-resolved vibrational spectroscopy detects protein-based intermediates in the photosynthetic oxygen-evolving cycle

Barry

Cooper²,

Riso³

et al. 2006

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

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Photosynthetic oxygen production by photosystem II (PSII) is responsible for the maintenance of aerobic life on earth. The production of oxygen occurs at the PSII oxygen-evolving complex (OEC), which contains a tetranuclear manganese (Mn) cluster. Photo-induced electron transfer events in the reaction center lead to the accumulation of oxidizing equivalents on the OEC. Four sequential photooxidation reactions are required for oxygen production. The oxidizing complex cycles among five oxidation states, called the S n states, where n refers to the number of oxidizing equivalents stored. Oxygen release occurs during the S 3-to-S0 transition from an unstable intermediate, known as the S4 state. In this report, we present data providing evidence for the production of an intermediate during each S state transition. These proteinderived intermediates are produced on the microsecond to millisecond time scale and are detected by time-resolved vibrational spectroscopy on the microsecond time scale. Our results suggest that a protein-derived conformational change or proton transfer reaction precedes Mn redox reactions during the S 2-to-S3 and S 3-to-S0 transitions.manganese cluster ͉ photosynthesis ͉ photosystem II ͉ time-resolved IR ͉ water oxidation T ime-resolved vibrational spectroscopy can detect chemical intermediates formed during enzymatic catalysis. Advantages include the technique's exquisite structural sensitivity and its high temporal resolution. Recent advances in methodology and interpretation have produced insights into the catalytic mechanism in several biological systems (for examples, see refs. 1-4).In this paper, we report the use of time-resolved IR spectroscopy to investigate the mechanism of photosynthetic water oxidation. Photosystem II (PSII) catalyzes the oxidation of water and the reduction of bound plastoquinone. Photoexcitation of PSII leads to the oxidation of the chlorophyll donor, P 680 , and the sequential reduction of a pheophytin (Fig. 1A, reaction 1) and a plastoquinone, Q A ( Fig. 1 A, reaction 2), in picoseconds. Q A reduces Q B to generate a semiquinone radical, Q B Ϫ , on the microsecond time scale (Fig. 1 A, reaction 3 ) (reviewed in ref. 5).A second photoexcitation leads to the reduction and protonation of Q B Ϫ to form the quinol Q B H 2 . The rate of reduction of Q B is faster than the rate of reduction of Q B Ϫ (see ref. 6 and references therein), which gives rise to a characteristic period-2 oscillation in kinetics originating on the PSII acceptor side (7).The primary chlorophyll donor, P 680 , oxidizes a tyrosine, Y Z (Y161 in the D1 subunit), on the nanosecond to microsecond time scale (Fig. 1 A, reaction 4). In turn, tyrosine Y Z ⅐ oxidizes the oxygen-evolving complex (OEC) on every flash (Fig. 1 A, reaction 5) (8). Four sequential photooxidation reactions are required for oxygen production, and the oxidizing complex cycles among five oxidation states, called the S n states, where n refers to the number of oxidizing equivalents stored (9). The rate of OEC oxidation slows as o...

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“…On the other hand gold can be used as an electrode for the electrochemical studies of the immobilized proteins [31], where the mediated electrical communication of an enzyme [25,[32][33][34] or antibody orientation [35] or an allosteric protein [19,28] with the gold electrode can be finely tuned through protein orientations on the SAM.…”

Section: Introductionmentioning

confidence: 99%

Direct reagentless detection of the affinity binding of recombinant His-tagged firefly luciferase with a nickel-modified gold electrode

Vagin

Trashin

Белоглазкина

et al. 2015

Mendeleev Communications

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The direct reagentless electrochemical detection of recombinant firefly luciferase binding with gold electrode modified with nickel ions has been carried out. Self-assembly of molecules of functional disulfide containing chelating moieties on the gold electrode was monitored by contact angle measurements and confirmed by electrochemical data as three times decrease of electric double layer capacitance. Succeeding nickel ions chelating followed by selective binding of target protein were registered by reagentless electrochemical measurements as consistent decreases of double layer capacitance. The quantification of bound protein has been achieved by means of luminescent assay of residual enzymatic activity in solution for electrode recovery. The electrode surface coverage by protein globules has been confirmed by AFM.

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Oriented Attachment and Membrane Reconstitution of His-Tagged CytochromecOxidase to a Gold Electrode: In Situ Monitoring by Surface-Enhanced Infrared Absorption Spectroscopy

Cited by 271 publications

References 45 publications

Resolving voltage-dependent structural changes of a membrane photoreceptor by surface-enhanced IR difference spectroscopy

Resolving voltage-dependent structural changes of a membrane photoreceptor by surface-enhanced IR difference spectroscopy

Time-resolved vibrational spectroscopy detects protein-based intermediates in the photosynthetic oxygen-evolving cycle

Direct reagentless detection of the affinity binding of recombinant His-tagged firefly luciferase with a nickel-modified gold electrode

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