Simultaneous Observation of Peptide Backbone Lipid Solvation and α‐Helical Structure by Deep‐UV Resonance Raman Spectroscopy

Halsey, Christopher M.; Xiong, Jian; Oshokoya, Olayinka O.; Johnson, J. A.; Shinde, Sandip S.; Beatty, J. Thomas; Ghirlanda, Giovanna; JiJi, Renee D.; Cooley, Jason W.

doi:10.1002/cbic.201100433

Cited by 22 publications

(30 citation statements)

References 28 publications

(10 reference statements)

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“…While the position of the amide I band (1652 cm −1 ) is consistent with α‐helical structure, the intensity of the amide I maximum is much stronger than typically observed for globular α‐helical proteins. The strong intensity of the amide I band indicates that the backbone of melittin is desolvated in the presence of DMPC bilayers consistent with previous dUVRR studies of lipid solvated proteins …”

Section: Resultssupporting

confidence: 89%

“…dUVRR spectra of proteins are dependent on their secondary structure composition. Recent studies have shown that dUVRR spectroscopy is amenable to structural studies of membrane‐solubilized proteins . There are four observable amide modes, the amide I, II, III, and S. The amide I is a carbonyl stretching mode and is typically quite weak in α‐helical and disordered conformations.…”

Section: Resultsmentioning

confidence: 99%

“…As the dUVRR spectra, like IR, represent the sum of all of a protein's secondary structural elements, the spectra can, therefore, be deconvoluted into the pure secondary structure content components . Only recently dUVRR spectroscopy has been applied to the field of membrane proteins revealing that selective excitation of the amide backbone results in a strong protein signal with little interference from the lipid and buffer components. Furthermore, the dUVRR spectra of lipid‐desolvated proteins have an enhanced amide I mode .…”

Section: Introductionmentioning

confidence: 99%

“…Only recently dUVRR spectroscopy has been applied to the field of membrane proteins revealing that selective excitation of the amide backbone results in a strong protein signal with little interference from the lipid and buffer components. Furthermore, the dUVRR spectra of lipid‐desolvated proteins have an enhanced amide I mode . Yet despite its promise as a complimentary tool for the study of membrane proteins, further work is needed to determine the effects of the membrane environment on the dUVRR spectra of proteins.…”

Section: Introductionmentioning

confidence: 99%

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Effects of fluidity on the ensemble structure of a membrane embedded α‐helical peptide

2014

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Melittin, the main hemolytic component of honeybee venom, is unfolded in an aqueous environment and folds into an α-helical conformation in a lipid environment. Membrane fluidity is known to affect the activity and structure of melittin. By combining two structurally sensitive optical methods, circular dichroism (CD) and deep-ultraviolet resonance Raman spectroscopy (dUVRR), we have identified distinct structural fluctuations in melittin correlated with increased and decreased 1,2-dimyristoyl-sn-glycero-3-phosphocholine bilayer fluidities. CD spectra have reduced intensity at temperatures above 22°C and high concentrations of the cholesterol analog 5α-cholestan-3β-ol indicating distortions in the α-helical structure under these conditions. No increase in the amide S is observed in the temperature-dependent dUVRR spectra, suggesting an increase in 310 -helical structure with increasing temperatures above 22°C. However, incorporation of 25 mol% 5α-cholestan-3β-ol resulted in a small increase in the amide S intensity indicating partial unfolding of melittin.

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Section: Resultssupporting

confidence: 89%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effects of fluidity on the ensemble structure of a membrane embedded α‐helical peptide

2014

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“…Comparison of the UVRR spectrum of this peptide to that of the soluble α -helical protein myoglobin led to the conclusion that the intensity of UVRR amide bands serves as a reliable marker to identify lipid-solubilized and solvent-exposed helical structure in proteins. 146 A final example of a membrane-associated peptide for folding studies is the α -helical, pH low insertion peptide (pHLIP). This peptide exhibits structural changes as a function of pH, and UVRR experiments determined that this peptide is desolvated and structured as a membrane-associated peptide.…”

Section: Ultraviolet Resonance Raman (Uvrr) Spectroscopymentioning

confidence: 99%

Insights into Protein Structure and Dynamics by Ultraviolet and Visible Resonance Raman Spectroscopy

et al. 2015

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Raman spectroscopy is a form of vibrational spectroscopy based on inelastic scattering of light. In resonance Raman spectroscopy, the wavelength of the incident light falls within an absorption band of a chromophore, and this overlap of excitation and absorption energy greatly enhances the Raman scattering efficiency of the absorbing species. The ability to probe vibrational spectra of select chromophores within a complex mixture of molecules makes resonance Raman spectroscopy an excellent tool for studies of biomolecules. In this Current Topic, we discuss the type of molecular insights obtained from steady-state and time-resolved resonance Raman studies of a prototypical photoactive protein, rhodopsin. We also review recent efforts in ultraviolet resonance Raman investigations of soluble and membrane-associated biomolecules, including integral membrane proteins and antimicrobial peptides. These examples illustrate that resonance Raman is a sensitive, selective, and practical method for studying the structures of biological molecules, and the molecular bonding, geometry, and environments of protein cofactors, the backbone, and side chains.

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Structures and Dynamics of Proteins Probed by UV Resonance Raman Spectroscopy

Leigh¹,

Schlamadinger²,

Kim³

2014

Biophysical Methods for Biotherapeutics

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Simultaneous Observation of Peptide Backbone Lipid Solvation and α‐Helical Structure by Deep‐UV Resonance Raman Spectroscopy

Cited by 22 publications

References 28 publications

Effects of fluidity on the ensemble structure of a membrane embedded α‐helical peptide

Effects of fluidity on the ensemble structure of a membrane embedded α‐helical peptide

Insights into Protein Structure and Dynamics by Ultraviolet and Visible Resonance Raman Spectroscopy

Structures and Dynamics of Proteins Probed by UV Resonance Raman Spectroscopy

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