title: Design and performance of an ultraviolet resonance Raman spectrometer for proteins and nucleic acids.
abstract: We describe an ultraviolet resonance Raman (UVRR) spectrometer appropriate for structural studies of biological macromolecules and their assemblies. Instrument design includes the following features: a continuous wave, intracavity doubled, ultraviolet laser source for excitation of the Raman spectrum; a rotating cell (or jet source) for presentation of the sample to the laser beam; a Cassegrain optic with f/1.0 aperture for collection of the Raman scattering; a quartz prism dispersing element for rejection of stray light and Rayleigh scattering; a 0.75-m single grating monochromator for dispersion of the Raman scattering; and a liquid-nitrogen-cooled, charge-coupled device for detection of the Raman photons. The performance of this instrument, assessed on the basis of the observed signal-to-noise ratios, the apparent resolution of closely spaced spectral bands, and the wide spectrometer bandpass of 2200 cm-1, is believed superior to previously described UVRR spectrometers of similar design. Performance characteristics of the instrument are demonstrated in UVRR spectra obtained from standard solvents, p-ethylphenol, which serves as a model for the tyrosine side chain, the DNA nucleotide deoxyguanosine-5'-monophosphate, and the human tumor necrosis factor binding protein, which is considered representative of soluble globular proteins.
authors.affiliation: Division of Cell Biology and Biophysics School of Biological Sciences, University of Missouri-Kansas City 64110-2499, USA.
authors.forename: M P
authors.initials: MP
authors.lastname: Russell
authors.forename: S
authors.initials: S
authors.lastname: Vohník
authors.forename: G J
authors.initials: GJ
authors.lastname: Thomas
mesh_terms: D000097:Acetonitriles / Q000737:chemistry; D055592:Biophysical Phenomena; D001703:Biophysics; D002251:Carbon Tetrachloride / Q000737:chemistry; D002352:Carrier Proteins / Q000737:chemistry; D003848:Deoxyguanine Nucleotides / Q000737:chemistry; D005069:Evaluation Studies as Topic; D006801:Humans; D046911:Macromolecular Substances; D008956:Models, Chemical; D015394:Molecular Structure; D009696:Nucleic Acids / Q000737:chemistry*; D010636:Phenols / Q000737:chemistry; D010777:Photochemistry; D011506:Proteins / Q000737:chemistry*; D018124:Receptors, Tumor Necrosis Factor; D047888:Receptors, Tumor Necrosis Factor, Type I; D013056:Spectrophotometry, Ultraviolet / Q000295:instrumentation*; D013059:Spectrum Analysis, Raman / Q000295:instrumentation*; D053319:Tumor Necrosis Factor Decoy Receptors; D014443:Tyrosine / Q000737:chemistry; D014466:Ultraviolet Rays
chemical_list: D000097:Acetonitriles; D002352:Carrier Proteins; D003848:Deoxyguanine Nucleotides; D046911:Macromolecular Substances; D009696:Nucleic Acids; D010636:Phenols; D011506:Proteins; D018124:Receptors, Tumor Necrosis Factor; D047888:Receptors, Tumor Necrosis Factor, Type I; D053319:Tumor Necrosis Factor Decoy Receptors; C069521:recombinant human tumor necrosis factor-binding protein-1; D014443:Tyrosine; C007257:2'-deoxyguanosine 5'-phosphate; C042291:4-ethylphenol; D002251:Carbon Tetrachloride; C032159:acetonitrile