Polarized raman scattering from oriented single microcrystals of d(A5T5)2 and d(pTpT)

Polarized Raman spectra have been obtained from single microcrystals of the duplex of the decamer d(A5T5)2 using a Raman microscope. This is the first report of Raman spectra from a crystal of a deoxyoligomer that contains only long, nonalternating sequences of adenine and thymine. Sequences contain...

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Veröffentlicht in:Biopolymers 1988-03, Vol.27 (3), p.493-507
Hauptverfasser: Patapoff, Thomas W., Thomas, Gerald A., Wang, Yang, Peticolas, Warner L.
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Sprache:eng
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Zusammenfassung:Polarized Raman spectra have been obtained from single microcrystals of the duplex of the decamer d(A5T5)2 using a Raman microscope. This is the first report of Raman spectra from a crystal of a deoxyoligomer that contains only long, nonalternating sequences of adenine and thymine. Sequences containing d(A)n and d(T)n are of interest in view of recent suggestions that they induce bends in DNA and that they might exist in a nonstandard B‐conformation. Polarized Raman spectra of a crystal of d(pTpT) have also been obtained. Both crystals display Raman bands whose intensities are very sensitive to the orientation of the crystal with respect to the direction of polarization of the incident laser beam. These spectra indicate that the helical axes of the oligonucleotides are parallel to the long axes of the crystals and that the d(A5T5)2 is not appreciably bent in the crystal. The Raman spectrum from the d(pTpT) crystal indicates that all of the furanose ring puckers are in a C2′‐endo configuration since only the C2′‐endo marker band at 835 ± 5 cm−1 is present. Crystals of d(A5T5)2 show measurable Raman intensities in both the 838‐ and 816‐cm−1 bands. This indicates the presence of both the C2′‐endo and C3′‐endo, or possibly other non‐C2′‐endo, furanose conformations. The 816‐cm−1 band is weak so that only a small fraction of the residues are estimated to be in the non‐C2′‐endo conformation. In both the d(pTpT) and d(A5T5)2 crystals the intensity of the bands due to vibrations of the backbone show only a small dependence on orientation of the crystals. This result is explained by the low symmetry of the puckered sugar rings. It is concluded that Raman spectra obtained from oligonucleotide crystals in which the orientation of the crystal axes to the laser polarization is not carefully controlled may contain intensity artifacts that are due to polarization effects.
ISSN:0006-3525
1097-0282
DOI:10.1002/bip.360270310