[17] Nanosecond time-resolved resonance Raman spectroscopy

Varotsis, Constantinos; Gt, Babcock

doi:10.1016/0076-6879(93)26019-6

Cited by 16 publications

(13 citation statements)

References 34 publications

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“…The flow-flash technique (Greenwood and Gibson, 1967), where CO is photodissociated in the presence of molecular oxygen, has delivered significant insights into the catalytic mechanism of CcO. This method has been adapted to resonance Raman spectroscopy (for review, see Varotsis and Babcock, 1993) where the involvement of the heme moiety could be elucidated on the vibrational level. Infrared absorption spectroscopy provides details beyond the heme modes to the vibrational changes of the entire apoprotein.…”

Section: Discussionmentioning

confidence: 99%

Transient Binding of CO to CuB in Cytochrome c Oxidase Is Dynamically Linked to Structural Changes around a Carboxyl Group: A Time-Resolved Step-Scan Fourier Transform Infrared Investigation

Heitbrink

Sigurdson

Bolwien

et al. 2002

Biophysical Journal

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The redox-driven proton pump cytochrome c oxidase is that enzymatic machinery of the respiratory chain that transfers electrons from cytochrome c to molecular oxygen and thereby splits molecular oxygen to form water. To investigate the reaction mechanism of cytochrome c oxidase on the single vibrational level, we used time-resolved step-scan Fourier transform infrared spectroscopy and studied the dynamics of the reduced enzyme after photodissociation of bound carbon monoxide across the mid-infrared range (2300-950 cm(-1)). Difference spectra of the bovine complex were obtained at -20 degrees C with 5 micros time resolution. The data demonstrate a dynamic link between the transient binding of CO to Cu(B) and changes in hydrogen bonding at the functionally important residue E(I-286). Variation of the pH revealed that the pK(a) of E(I-286) is >9.3 in the fully reduced CO-bound oxidase. Difference spectra of cytochrome c oxidase from beef heart are compared with those of the oxidase isolated from Rhodobacter sphaeroides. The bacterial enzyme does not show the environmental change in the vicinity of E(I-286) upon CO dissociation. The characteristic band shape appears, however, in redox-induced difference spectra of the bacterial enzyme but is absent in redox-induced difference spectra of mammalian enzyme. In conclusion, it is demonstrated that the dynamics of a large protein complex such as cytochrome c oxidase can be resolved on the single vibrational level with microsecond Fourier transform infrared spectroscopy. The applied methodology provides the basis for future investigations of the physiological reaction steps of this important enzyme.

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Section: Discussionmentioning

confidence: 99%

Transient Binding of CO to CuB in Cytochrome c Oxidase Is Dynamically Linked to Structural Changes around a Carboxyl Group: A Time-Resolved Step-Scan Fourier Transform Infrared Investigation

Heitbrink

Sigurdson

Bolwien

et al. 2002

Biophysical Journal

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“…After photolysis of MbCO, TR 3 spectroscopy at this resonance wavelength shows two distinct conformations of the heme pocket that are associated with different Fe(II)-CO vibrational frequencies (64). The combination of selectivity and structural sensitivity makes resonance Raman spectroscopy a powerful tool for structural studies (see 18,42,73).…”

Section: Time-resolved Resonance Raman (Tr 3 ) Spectroscopymentioning

confidence: 98%

Nanosecond Time-Resolved Spectroscopy of Biomolecular Processes

Chen

Goldbeck²,

Kliger³

1997

Annu. Rev. Biophys. Biomol. Struct.

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Over the past two decades, nanosecond absorption and vibrational spectroscopies have developed into powerful tools for monitoring the secondary, tertiary, and quaternary structural relaxations of biological macromolecules under near-physiological conditions of solvent and temperature. Observed through such methods, the dynamic response of a biomolecule to photoinitiated excursions from equilibrium can reveal valuable information about the structure-function relationship, information beyond that obtained from the static structures provided by X-ray crystallography, nuclear magnetic resonance spectroscopy, and other steady-state methods. Most recently, the development of ultra-sensitive polarization techniques for absorption spectroscopy has greatly enhanced the amount of time-resolved structural information that can be obtained from the broadened electronic spectra of biomolecules. This review examines nanosecond absorption, vibrational, and polarized absorption methods, and their applications to protein function and folding, emphasizing the complementary nature of information obtained from electronic and vibrational spectra measured on the nanosecond time scale.

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“…In most of the studies reviewed in the following sections, human adult Hb (HbA) was employed. Early studies up to the midnineties were reviewed by Babcock and Kincaid, with special focus on the experimental approaches. , …”

Section: Ligand Bindingmentioning

confidence: 99%

“…Early studies up to the midnineties were reviewed by Babcock and Kincaid, with special focus on the experimental approaches. 305,306 5.1.2. Ligand Dissociation and Rebinding.…”

Section: Hemoglobinmentioning

confidence: 99%

“…The various experimental TR RR spectroscopic approaches for studying CcO were described in previous reviews. 305,306,463 In general, the concept utilizes the fact that the reduced BNC can bind CO. 464 If, after mixing with an O 2 -saturated buffer solution, the CcO− CO complex is photolyzed immediately, the high binding rate of molecular oxygen ensures the formation of the CcO-O 2 complex rather than CO rebinding. 465 CcO-O 2 formation constitutes the starting point for the subsequent catalytic reaction cycle including intermediates with lifetimes on the μs and ms time scale.…”

Section: Cytochrome C Oxidasementioning

confidence: 99%

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Probing Structure and Reaction Dynamics of Proteins Using Time-Resolved Resonance Raman Spectroscopy

Buhrke

Hildebrandt

2019

Chem. Rev.

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The mechanistic understanding of protein functions requires insight into the structural and reaction dynamics. To elucidate these processes, a variety of experimental approaches are employed. Among them, time-resolved (TR) resonance Raman (RR) is a particularly versatile tool to probe processes of proteins harboring cofactors with electronic transitions in the visible range, such as retinal or heme proteins. TR RR spectroscopy offers the advantage of simultaneously providing molecular structure and kinetic information. The various TR RR spectroscopic methods can cover a wide dynamic range down to the femtosecond time regime and have been employed in monitoring photoinduced reaction cascades, ligand binding and dissociation, electron transfer, enzymatic reactions, and protein un-and refolding. In this account, we review the achievements of TR RR spectroscopy of nearly 50 years of research in this field, which also illustrates how the role of TR RR spectroscopy in molecular life science has changed from the beginning until now. We outline the various methodological approaches and developments and point out current limitations and potential perspectives.

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[17] Nanosecond time-resolved resonance Raman spectroscopy

Cited by 16 publications

References 34 publications

Transient Binding of CO to CuB in Cytochrome c Oxidase Is Dynamically Linked to Structural Changes around a Carboxyl Group: A Time-Resolved Step-Scan Fourier Transform Infrared Investigation

Transient Binding of CO to CuB in Cytochrome c Oxidase Is Dynamically Linked to Structural Changes around a Carboxyl Group: A Time-Resolved Step-Scan Fourier Transform Infrared Investigation

Nanosecond Time-Resolved Spectroscopy of Biomolecular Processes

Probing Structure and Reaction Dynamics of Proteins Using Time-Resolved Resonance Raman Spectroscopy

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