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Biomolecular ligands screening using radiation damping difference WaterLOGSYspectroscopy

Supplementary Material

Peng Sun1,2, Xianwang Jiang1,2, Bin Jiang1, Xu Zhang1*, Maili Liu1* 1. Wuhan Center for Magnetic Resonance, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan 430071 2. Graduate School of Chinese Academy of Sciences, Beijing 100049

1 Fig. S1 800 MHz spectra of RD-WaterLOGSY (a), PO-WaterLOGSY (b), and WaterLOGSY, all with water flip-back as read pulse, double W5 for water suppression and mixing time of 1.4 s. The sample contains 0.1 mM HSA as receptor protein, 2.0 mM Trp and 2.0 mM Glu as binding and non-binding ligands, respectively. The spectra were recorded at Bruker Avance 800 spectrometer with a cryoprobe. The number of scans, dummy scans, and the number of data points in the time domain were 32, 8, and 65536, respectively. The recovery delay D1 of 2.5 s was used for all experiment with exception of 0.0 s for the odd scans of PO-WaterLOGSY.

2 Pulse sequence code of radiation damping difference (RDD) WaterLOGSY on Bruker Avance spectrometer (topspin 2.0)

; RDD WaterLOGSY ; For biomolecule ligands screening at natural abundance ; Written by Peng Sun 2011/10/11 ; See footnotes

;avance-version (07/01/22) ;1D sequence

;$CLASS=HighRes ;$DIM=1D ;$TYPE= ;$SUBTYPE= ;$COMMENT=

#include #include #include

"p2=p1*2"

"DELTA1=d19-p27*0.174/3.1416" "DELTA2=d19-p27*0.412/3.1416" "DELTA3=d19-p27*0.826/3.1416" "DELTA4=d19-p27*1.556/3.1416" "DELTA5=d19-p27*2.982/3.1416" "DELTA6=d8-d16*2-p17-20u"

1 ze 2 30m 20u

3 d1 pl1:f1 50u UNBLKGRAD p2 ph1 vd p17:gp3 d16 gron0 6u ivd vd DELTA6 10u groff d16

(p11:sp1 ph6:r):f1 4u pl1:f1 p1 ph2

50u pl18:f1 p16:gp1 d16 p27*0.087 ph4 DELTA1 DELTA2 p27*0.206 ph4 DELTA2 DELTA3 p27*0.413 ph4 DELTA3 DELTA4 p27*0.778 ph4 DELTA4 DELTA5 p27*1.491 ph4

4 DELTA5 DELTA5 p27*1.491 ph5 DELTA5 DELTA4 p27*0.778 ph5 DELTA4 DELTA3 p27*0.413 ph5 DELTA3 DELTA2 p27*0.206 ph5 DELTA2 DELTA1 p27*0.087 ph5 50u p16:gp1 d16

4u p16:gp2 d16 p27*0.087 ph4 DELTA1 DELTA2 p27*0.206 ph4 DELTA2 DELTA3 p27*0.413 ph4 DELTA3 DELTA4 p27*0.778 ph4 DELTA4 5 DELTA5 p27*1.491 ph4 DELTA5 DELTA5 p27*1.491 ph5 DELTA5 DELTA4 p27*0.778 ph5 DELTA4 DELTA3 p27*0.413 ph5 DELTA3 DELTA2 p27*0.206 ph5 DELTA2 DELTA1 p27*0.087 ph5 p16:gp2 d16

4u BLKGRAD go=2 ph31 30m mc #0 to 2 F0(zd) exit ph1=0 ph2=0 0 2 2 ph4=0 ph5=2 ph6=0 2 2 0 ph31=0 2 2 0

6 ;------NOTES------;NS=4*n ;DS≥8

; 1H pulses ;p1: 90 degree high power 1H pulse @pl1 ;p27: 90 degree high power1H pulse @pl18 ;p11: 90 degree 1H shape pulse ;pl1: power level for 1H 90 degree ;pl18: power level for 1H 90 degree for WATERGATE W5; ;sp11: power level for p11

; delay ;d8: mixing time [1-2.5s] ;d16: delay for homospoil/gradient recovery. ;d19: delay for binomial water suppression ; d19 = (1/(2*d)), d = distance of next null (in Hz)

;vd: variable delay for the control of radiation damping, taken from vd-list user defined.[4u, 12ms].

; gradients ;p16: 1000u ;p30: 3000u

;for z-only gradients: ;gpz1: 34% ;gpz2: 22% ;gpz3: 75% ;gpz4: 0.1%

;use gradient files: ;gpnam1: SINE.100 ;gpnam2: SINE.100 ;gpnam3: SINE.100 7 8