Raman spectroscopy in bioanalysis

Pappas, Dimitri; Smith, Benjamin W.; Winefordner, J. D.

doi:10.1016/s0039-9140(99)00254-4

Cited by 99 publications

(67 citation statements)

References 64 publications

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“…The advent of the holographic notch filter to reject more effectively elastically scattered (Rayleigh) light, introduction of the volumephase holographic transmissive grating to maximize photon throughput and implementation of efficient charge-coupleddevice (CCD) detectors also have contributed to dramatic increases in spectrophotometric sensitivity and selectivity. 54 Transmissive optics also provide an appreciably more stable Raman platform that preserves spectrometer calibration and alignment. 55 This is of particular importance in the application of Raman digital difference spectroscopy, a very useful and powerful approach for characterizing conformational changes in nucleic acid complexes and other biomolecular assemblies.…”

Section: Overviewmentioning

confidence: 99%

Raman, polarized Raman and ultraviolet resonance Raman spectroscopy of nucleic acids and their complexes

Benevides

Overman

Thomas

2005

J Raman Spectroscopy

200

231

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Applications of Raman spectroscopy to investigate the molecular constituents of nucleic acids were initiated in the late 1960s and soon thereafter progressed to studies of synthetic and native nucleic acids and complex biological assemblies containing either DNA or RNA. Raman applications to nucleic acids have continued to increase in number and diversity up to the present time. This paper attempts to provide an overview of this large body of work, with emphasis on studies carried out during the past decade and focusing on problems of biological interest and significance. The specific Raman methodologies included in this review of nucleic acid applications are (i) conventional Raman spectroscopy (i.e. off-resonance Raman excitation), (ii) ultraviolet resonance Raman (UVRR) spectroscopy, and (iii) polarized Raman microspectroscopy. For each methodology, the experimentally obtained nucleic acid spectrum consists of a number of discrete vibrational bands, most of which can be assigned confidently to a base, sugar or phosphate constituent of the macromolecule and many of which can be employed as sensitive indicators or fingerprints of either local structure, global conformation, intermolecular interaction or molecular dynamics. The applications selected for review include numerous examples from the authors' laboratories. The topics addressed include the influences of base composition, base sequence, superhelical stress and drug ligation on nucleic acid structure and polymorphism, the thermodynamic parameters governing nucleic acid premelting and melting phenomena, the molecular mechanisms and determinants of protein/nucleic acid recognition and the structures and dynamics of nucleic acids in virus assemblies.

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Section: Overviewmentioning

confidence: 99%

Raman, polarized Raman and ultraviolet resonance Raman spectroscopy of nucleic acids and their complexes

Benevides

Overman

Thomas

2005

J Raman Spectroscopy

200

231

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“…Accordingly, they may denature and the denatured protein may be different in the secondary structure or conformational integrity compared to that of the native protein. FT-Raman spectroscopy is a powerful tool for the characterisation of protein structure in diverse physicochemical environments (Pappas et al, 2000). For example, Quinn et al (1999) studied lysozyme structural integrity in the hydrofluoroalkane propellants tetrafluoroethane and heptafluoropropane by FT-Raman spectroscopy; also Kudryavtsev et al (2000) demonstrated the effect of ordering of internal water in thaumatin and lysozyme crystals by FT-Raman spectroscopy.…”

Section: Introductionmentioning

confidence: 99%

Study of protein conformational stability and integrity using calorimetry and FT-Raman spectroscopy correlated with enzymatic activity

Elkordy

Forbes²,

Barry³

2008

European Journal of Pharmaceutical Sciences

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Enzymatic assay after cooling of thermally denatured protein solutions from HSDSC determined if thermal transition reversibility was related to biological activity. HSDSC data showed that molecules from lyophilised lysozyme were able to refold better than the spraydried form. This was confirmed by enzymatic assay. Moreover, enzymatic assay results revealed that lysozyme folding reversibility was related to the native structure of the protein that is essential for the biological activity.Thermal denaturation of DNase I and LDH samples in HSDSC was not reversible upon cooling of thermally denatured proteins (in contrast to lysozyme). Hence, it was decided to identify the effect of protein initial structures on its propensity to thermal denaturation via FT-Raman spectroscopy. In other words, proteins may denature with structural alterations due to stresses such as heat and the protein loses its enzymatic activity. Consequently, FTRaman investigated the effects of spray drying and heating of solid DNase I and LDH samples, from differential scanning calorimetry, on protein conformational integrities. Lyophilised and spray-dried DNase I and LDH solid samples were heated to two temperatures, one before the apparent denaturation temperatures (T m ) and the other after the T m . Samples heated below their T m showed some alterations of the secondary structure and some enzy- IntroductionProteins fold in three dimensions. Protein structure is organised from primary structure to quaternary structure. primary structure or the amino acid sequence of the peptide chains is the ultimate determinant for the protein native form. The protein secondary structure is the assignment of ␣-helices and ␤-sheets along the main peptide chain; whilst the ter-0928-0987/$ -see front matter

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“…blood analysis), guiding of surgical interventions, and intraoperative tumour border determination. Since the differentiation of tissue is possible, it is predicted that Raman spectroscopy may soon become a valuable tool to assist in clinical pathology (17).…”

Section: Raman Spectroscopymentioning

confidence: 99%