caa a22 4500 445325348 CHVBK 20180317142718.0 cr unu---uuuuu 170323e20111201xx s 000 0 eng 10.1007/s10858-011-9568-3 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9568-3 Alternative SAIL-Trp for robust aromatic signal assignment and determination of the χ2 conformation by intra-residue NOEs [Elektronische Daten] [Yohei Miyanoiri, Mitsuhiro Takeda, JunGoo Jee, Akira Ono, Kosuke Okuma, Tsutomu Terauchi, Masatsune Kainosho] Tryptophan (Trp) residues are frequently found in the hydrophobic cores of proteins, and therefore, their side-chain conformations, especially the precise locations of the bulky indole rings, are critical for determining structures by NMR. However, when analyzing [U-13C,15N]-proteins, the observation and assignment of the ring signals are often hampered by excessive overlaps and tight spin couplings. These difficulties have been greatly alleviated by using stereo-array isotope labeled (SAIL) proteins, which are composed of isotope-labeled amino acids optimized for unambiguous side-chain NMR assignment, exclusively through the 13C-13C and 13C-1H spin coupling networks (Kainosho et al. in Nature 440:52-57, 2006). In this paper, we propose an alternative type of SAIL-Trp with the [ζ2,ζ3-2H2; δ1,ε3,η2-13C3; ε1-15N]-indole ring ([12C γ, 12 Cε2] SAIL-Trp), which provides a more robust way to correlate the 1Hβ, 1Hα, and 1HN to the 1Hδ1 and 1Hε3 through the intra-residue NOEs. The assignment of the 1Hδ1/13Cδ1 and 1Hε3/13Cε3 signals can thus be transferred to the 1Hε1/15Nε1 and 1Hη2/13Cη2 signals, as with the previous type of SAIL-Trp, which has an extra 13C at the Cγ of the ring. By taking advantage of the stereospecific deuteration of one of the prochiral β-methylene protons, which was 1Hβ2 in this experiment, one can determine the side-chain conformation of the Trp residue including the χ2 angle, which is especially important for Trp residues, as they can adopt three preferred conformations. We demonstrated the usefulness of [12Cγ,12Cε2] SAIL-Trp for the 12kDa DNA binding domain of mouse c-Myb protein (Myb-R2R3), which contains six Trp residues. Springer Science+Business Media B.V., 2011 SAIL-Trp nationallicence Aromatic ring CH assignment nationallicence Intra-residue NOE nationallicence χ2 conformation nationallicence Miyanoiri Yohei Graduate School of Science, Structural Biology Research Center, Nagoya University, Furo-cho, Chikusa-ku, 464-8602, Nagoya, Japan aut Takeda Mitsuhiro Graduate School of Science, Structural Biology Research Center, Nagoya University, Furo-cho, Chikusa-ku, 464-8602, Nagoya, Japan aut Jee JunGoo Center for Priority Areas, Tokyo Metropolitan University, 1-1 Minami-ohsawa, 192-0397, Hachioji, Japan aut Ono Akira Center for Priority Areas, Tokyo Metropolitan University, 1-1 Minami-ohsawa, 192-0397, Hachioji, Japan aut Okuma Kosuke Center for Priority Areas, Tokyo Metropolitan University, 1-1 Minami-ohsawa, 192-0397, Hachioji, Japan aut Terauchi Tsutomu Center for Priority Areas, Tokyo Metropolitan University, 1-1 Minami-ohsawa, 192-0397, Hachioji, Japan aut Kainosho Masatsune Graduate School of Science, Structural Biology Research Center, Nagoya University, Furo-cho, Chikusa-ku, 464-8602, Nagoya, Japan aut Journal of Biomolecular NMR Springer Netherlands 51/4(2011-12-01), 425-435 0925-2738 51:4<425 2011 51 10858 https://doi.org/10.1007/s10858-011-9568-3 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9568-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Miyanoiri Yohei Graduate School of Science, Structural Biology Research Center, Nagoya University, Furo-cho, Chikusa-ku, 464-8602, Nagoya, Japan aut NATIONALLICENCE 700 1- Takeda Mitsuhiro Graduate School of Science, Structural Biology Research Center, Nagoya University, Furo-cho, Chikusa-ku, 464-8602, Nagoya, Japan aut NATIONALLICENCE 700 1- Jee JunGoo Center for Priority Areas, Tokyo Metropolitan University, 1-1 Minami-ohsawa, 192-0397, Hachioji, Japan aut NATIONALLICENCE 700 1- Ono Akira Center for Priority Areas, Tokyo Metropolitan University, 1-1 Minami-ohsawa, 192-0397, Hachioji, Japan aut NATIONALLICENCE 700 1- Okuma Kosuke Center for Priority Areas, Tokyo Metropolitan University, 1-1 Minami-ohsawa, 192-0397, Hachioji, Japan aut NATIONALLICENCE 700 1- Terauchi Tsutomu Center for Priority Areas, Tokyo Metropolitan University, 1-1 Minami-ohsawa, 192-0397, Hachioji, Japan aut NATIONALLICENCE 700 1- Kainosho Masatsune Graduate School of Science, Structural Biology Research Center, Nagoya University, Furo-cho, Chikusa-ku, 464-8602, Nagoya, Japan aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 51/4(2011-12-01), 425-435 0925-2738 51:4<425 2011 51 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer