Structural studies of bean pod mottle virus, capsid, and RNA in crystal and solution states by laser Raman spectroscopy.
Structures of protein and RNA components of bean pod mottle virus (BPMV) have been investigated by use of laser Raman spectroscopy. Raman spectra were collected from both aqueous solutions and single crystals of BPMV capsids (top component) and virions (middle and bottom components, which package, respectively, small and large RNA molecules). Analysis of the data permits the assignment of conformation-sensitive Raman bands to viral protein and RNA constituents and observation of structural similarities and differences between solution and crystalline states of BPMV components. The Raman results show that the protein subunits of the empty capsid contain between 45% and 55% beta-strand and beta-turn secondary structure, in agreement with the recently determined X-ray crystal structure, and that this total beta-strand content undergoes a small increase (approximately 5%) with packaging of RNA. The subunits are relatively deficient in alpha-helix secondary structure, estimated at less than 25%, and therefore must contain extensive amounts (greater than 20%) of loops and irregular chain conformations. The Raman spectra also show the following: (1) The molecular environments of as many as four tryptophan residues per subunit are altered upon packaging RNA, resulting in stronger 1N-H hydrogen bonding for two Trp residues and more hydrophobic environments for two indole rings. (2) Hydrogen-bonding states of the seven Tyr residues per subunit do not change detectably when RNA is packaged. At least five tyrosine OH groups are involved exclusively as strong hydrogen bond donors to protein acceptor groups, which suggests restricted access of solvent H2O molecules to these parahydroxyls.(ABSTRACT TRUNCATED AT 250 WORDS)
Li
T S
TS
Division of Cell Biology and Biophysics, School of Basic Life Sciences, University of Missouri, Kansas City 64110-2499.
Chen
Z G
ZG
Johnson
J E
JE
Thomas
G J
GJ
D000596:Amino Acids / Q000235:genetics; D002213:Capsid; D004267:DNA Viruses / Q000235:genetics; D007887:Fabaceae / Q000382:microbiology; D009690:Nucleic Acid Conformation; D010942:Plant Viruses / Q000235:genetics*; D010946:Plants, Medicinal; D011487:Protein Conformation; D012367:RNA, Viral / Q000032:analysis*; D012996:Solutions; D013059:Spectrum Analysis, Raman; D014961:X-Ray Diffraction
D000596:Amino Acids; D012367:RNA, Viral; D012996:Solutions