J. Biochem, 2004, Vol. 136, No. 1 29-37
© 2004 The Japanese Biochemical Society
BIOCHEMISTRY |
Side Chain–Side Chain Interactions of Arginine with Tyrosine and Aspartic Acid in Arg/Gly/Tyr–Rich Domains within Plant Glycine-Rich RNA Binding Proteins
1 High-Resolution NMR Laboratory and 2 Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060-0810; and 3 School of Health Sciences, Sapporo Medical University, Sapporo 060-8556
Plant glycine-rich RNA-binding proteins (GRRBPs) contain a glycine-rich region at the C-terminus whose structure is quite unknown. The C-terminal glycine-rich part is interposed with arginine and tyrosine (arginine/glycine/tyrosine (RGY)–rich domain). Comparative sequence analysis of forty-one GRRBPs revealed that the RGY-rich domain contains multiple repeats of Tyr-(Xaa)h-(Arg)k-(Xaa)l, where Xaa is mainly Gly, "k" is 1 or 2, and "h" and "l" range from 0 to 10. Two peptides, 1 (G1G2Y3G4G5G6R7R8D9G10) and 2 (G1G2R3R4D5G6G7Y8G9G10), corresponding to sections of the RGY-rich domain in Zea mays RAB15, were selected for CD and NMR experiments. The CD spectra indicate a unique, positive band near 228 nm in both peptides that has been ascribed to tyrosine residues in ordered structures. The pH titration by NMR revealed that a side chain-side chain interaction, presumably an H-N
···O=C
hydrogen bonding interaction in the salt bridge, occurs between Arg (i) and Asp (i + 2). 1D GOESY experiments indicated the presence of NOE between the aromatic side chain proton and the arginine side chain proton in the two peptides suggesting strongly that the Arg (i) aromatic side chain interacts directly with the Tyr (i ± 4 or i ± 5) side chain.
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