Supplemental Data

Files in this Data Supplement:

• Supplemental Table 1 - Amino acid sequences, primary citations, and database access codes (SwissProt) for all cyclotides and related circular trypsin inhibitors published to date.
• Supplemental Table 2 - Chemical shift assignments of vhr1 in 70% H₂O/10% D₂O/20% acetonitrile-d₃ at 298 K and 750 MHz.
• References
• Supplemental Figure 1 - Fingerprint region of the NOESY spectrum (upper panel) and amide region of the TOCSY spectrum (lower panel) of vhr1 recorded at 310 K and 750 MHz. In the NOESY spectrum, Hα_i-HN_i+1 connectivities are shown by lines and intraresidue HN-Hα cross peaks are labeled with the residue number. The asterisks denote NOESY cross peaks of the two proline residues, Hα₉-HN₁₀ and Hα₃₀-HN₁. Squares in the upper panel indicate the positions of interresidual (open squares) and intraresidual (for C25, black square) NOE cross peaks not observed in this spectrum, but detectable in the dataset recorded at 298 K. In the TOCSY spectrum (lower panel), the spin systems are labeled with one letter code and residue number.
• Supplemental Figure 2 - Summary of the NMR data used to characterize the secondary structure of vhr1. The top of the diagram shows the loop numbering and amino acid sequence of the peptide. The Hα secondary chemical shift values (SCS, in ppm) at 298 K, i.e. the differences between observed and random coil chemical shifts, are followed by bars indicating sequential connectivities observed in a NOESY spectrum with a mixing time of 250 ms. The height of the bars corresponds to the intensity of the observed NOEs; open bars indicate Hα-Hδ_i+1 connectivities for the residues preceding P9 and P30. For coupling constants, up arrow indicates ³J_HN-Hα coupling constants >= 8 Hz, down arrow indicates ³J_HN-Hα coupling constants <= 5 Hz. The lines below indicate medium and long-range NOE connectivities, with the thickness of the line corresponding to the intensity of the observed NOE. To emphasize the circular nature of vhr1, the first three residues of the peptide are shown again in light gray at the end of the sequence. The residues are numbered at the bottom of the figure; the cysteine connectivities are indicated by the lines. Random coil chemical shift values were taken from Merutka et al. (1995), except for Gly Hα shifts, which were taken from Wishart et al. (1992).