/1.95 two.48/2.51 2.76/2.98 2.58/2.88 two.63/2.49 two.33/2.65 2.41/2.87 two.64/2.80 2.54/2.52 two.25/2.53 two.30/2.56 2.36/2.51 two.21/2.39 2.77/3.01 2.67/2.89 2.60/2.53 2.33/2.67 2.35/2.81 two.62 two.34/2.72 2.26/2.36 1.97/2.H30 4.3 four.75 4.97 four.96 five.03 four.96 five.10 5.00 four.91 four.77 four.81 four.85 4.86 4.96 four.95 5.03 5.03 5.09 five.00 4.87 four.73 four.H40 4.23 4.44 four.40 4.49 four.56 4.51 four.40 4.47 four.41 four.41 4.33 four.32 4.20 4.39 four.51 four.58 4.56 four.40 4.45 4.42 4.43 three.H50 /H500 3.37/3.77 3.78/4.07 4.03/4.24 four.23/4.26 four.36/4.22 4.22/4.28 four.18/4.28 4.24/4.31 4.15/4.27 3.99/4.13 four.05/4.11 four.10/4.24 three.93/4.01 four.02/4.27 four.16/4.28 four.22/4.34 four.22/4.30 four.19/4.28 4.13/4.19 four.13/4.20 four.12/4.19 three.66/3.TMe H six H1 ‘ H2’, ” H3’TH6 Me

/1.95 2.48/2.51 2.76/2.98 2.58/2.88 two.63/2.49 two.33/2.65 two.41/2.87 2.64/2.80 2.54/2.52 two.25/2.53 two.30/2.56 two.36/2.51 2.21/2.39 two.77/3.01 2.67/2.89 2.60/2.53 two.33/2.67 two.35/2.81 two.62 2.34/2.72 two.26/2.36 1.97/2.H30 4.three four.75 4.97 4.96 5.03 4.96 5.ten 5.00 4.91 4.77 four.81 4.85 four.86 four.96 4.95 five.03 five.03 five.09 five.00 four.87 4.73 four.H40 4.23 4.44 4.40 four.49 4.56 4.51 four.40 four.47 4.41 four.41 four.33 4.32 four.20 four.39 four.51 four.58 four.56 four.40 four.45 4.42 4.43 3.H50 /H500 3.37/3.77 three.78/4.07 four.03/4.24 4.23/4.26 4.36/4.22 4.22/4.28 four.18/4.28 four.24/4.31 four.15/4.27 three.99/4.13 four.05/4.11 4.10/4.24 three.93/4.01 four.02/4.27 four.16/4.28 4.22/4.34 4.22/4.30 four.19/4.28 4.13/4.19 four.13/4.20 four.12/4.19 three.66/3.TMe H six H1 ‘ H2′, ” H3’TH6 Me H1′ H2′ H2” H3′ H4’H8H1’H2’H2”H3’H4′ H5′,” H7.24 7.57 8.00 7.67 7.72 7.91 7.97 7.94 7.78 7.85 7.86 7.71 7.51 eight.13 7.77 7.75 7.94 7.96 7.9 7.42 7.63 7.five.55 ten.65b 11.69 11.19 11.01 6.09 11.64 11.49 11.29 six.10 6.08 1.87 1.57 11.94 11.26 11.03 six.11 11.16 11.32 10.97 ten.20b 1.five.67 5.33 six.06 6.11 six.36 6.39 6.08 6.05 six.34 six.26 six.27 6.28 5.99 6.08 6.16 six.39 six.42 six.07 five.93 5.97 5.6 five.H8H1’H2’H2”H3′ H4′ H5′,” HGH8H1’H2’H2”H3’H4′ H5′,” H1 GG14 H1H1’H2’H2”H3’H4’H5′,”HH1H1’H2’H2”H3′ H4′ H5′,”HGCH3′ H2′ H2” H1′ HGH8H1′ H2′ H2” H3′ H4′ H5′ ” H,H8H1’H2’H2”H3′ H4′ H5′,” H1 GGH1H1’H2’H2” H3’H4’H5”HH1 H1’H2’H2”H3’H4’H5′,”HGGH8H1’H2’H2”H3’H4′ H5′ ” H,H8H1’H2’H2”H3’H4′ H5′,” H1 GCH 1′ H 2′ H 2’ ‘HG16 H1H1’H2’H2” H3′ H4′ H5′,”H3’H1 H1’H2’H2”H3’H4′ H5′,”H,GH8 H1’H2’H2”H3’H4’H5′ ” HHG21 TH6 H1’H2’H2”H3’H4’H5’,”MeFigure four. Schematic diagram of inter-residue NOE connectivities of Pu22-T12T13.bThe chemical shifts are measured in 25 mM K- phosphate, 70 mM KCl (pH 7.0) referenced to DSS. Chemical shift measured at 2 C.side. Sequential NOEs for stacking interactions are not observed for the other 3 residues in the 4-nt loop, and clearly downfield-shifted chemical shifts are suggestive of their groove place.H2N-PEG2-CH2COOtBu Order A number of NOEs are observed for the two flanking sequences, both of which seem to adopt well-stacking conformations.866641-66-9 Formula For the 50 -flanking C1-G2 segment, sequential NOEs are observed at the G2-G3 step, for example G3H8/G2H8, G3H8/G2H10 , H20 ,00 and H30 , at the same time as in the C1-G2 step (Figures 3B and 4).PMID:23341580 Surprisingly, the NOE between G2H8/G18H1 was powerful, indicating that G2 stacks totally above the 50 -tetrad with its H8 end positioned proper above G18H1. Similar sequential NOEs are observed for the 30 -flanking G21-T22 segment, i.e. at G21-G20 and T22-G21 measures (Figures 3B and four). A clear NOE observed amongst G21H8/G20H1 (Figure 3A) indicates that G21 stacks nicely with G20. NOE-refined solution structure with the VEGF G-quadruplex shows unique capping structure involving each the 4-nt middle loop and also the two flanking segments Resolution structures with the Pu22-T12T13 G-quadruplex have been calculated using a NOE-restrained distance geometry (DGSA) and restrained molecular dynamics (RMD) approach (Figure 5, PDB ID 2m27), beginning from an arbitrary extended single-stranded DNA. A total of 407 NOE distance restraints, such as 145 interresidue NOE interactions, have been utilized in the NOE-restrained structure calculation (Supplementary Table S1). Dihedral restraints are made use of for the anti glycosidic torsion angle () for loop residues. The stereo view with the ten lowest power structures is shown in Figure 5A. Thestructure statistics are listed in Supplementary Table S1. Pu22-T12T13 forms a well-defined parallel-stranded Gquadruplex structure with 3 tetrads. The two 1.