Supplemental Material for
Mapping of neuropeptides in the crustacean stomatogastric
nervous system by imaging mass spectrometry
Hui Ye1†, Limei Hui2†, Katherine Kellersberger3 and Lingjun Li1,2,*
1
School of Pharmacy and 2Department of Chemistry, University of Wisconsin-Madison, 777
Highland Avenue, Madison, WI 53705-2222, USA
3
Bruker Daltonics Inc., 40 Manning Rd, Billerica, MA 01821-3915, USA
†
The authors contributed equally to the manuscript.
1
Supplemental Table 1. Peptides identified from C. sapidus STG by MALDI-TOF/TOF,
MALDI-FTICR and ESI-Q-TOF MS platforms.
m/z
AST-A
909.4941
937.4890
962.5094
AST-B
1107.5153
1123.5147
1165.5538
1182.5691
1209.5800
1220.5807
1222.5752
1246.6076
1252.5858
1293.6335
1470.7025
Orcokinin
1062.5578
1156.5269
1186.5375
1198.5487
1228.5593
1256.5542
1286.5648
1403.6226
1433.6332
1474.6597
1502.6910
1504.6703
1532.7016
1547.6761
Sequence
TOF/TOF
FTICR
ARPYSFGLa
PRDYAFGLa
APQPYAFGLa
+
+
-
+
-
CoG/stn
CoG/stn
stn
AGWSSMRGAWa
AGWSSM(O)RGAWa
NWNKFQGSWa
TSWGKFQGSWa
TGWNKFQGSWa
SGDWSSLRGAWa
GNWNKFQGSWa
GSNWSNLRGAWa
NNWSKFQGSWa
STNWSSLRSAWa
VPNDWAHFRGSWa
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
CoG/stn
TPRDIANLYa
FDEIDRSGFA
FDEIDRSSFA
NFDEIDRSGFa
NFDEIDRSSFa
NFDEIDRSGFG
NFDEIDRSSFG
NFDEIDRSGFGF
NFDEIDRSSFGF
NFDEIDRSGFGFA
NFDEIDRSGFGFV
NFDEIDRSSFGFA
NFDEIDRSSFGFV
NFDEIDRSSFGFN
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
-
+
+
+
+
+
+
+
+
2
Q-TOF
Other
locationsa
stn
stn
stn
stn
stn
CoG/stn
CoG/stn/OG
CoG/stn
OG
Stn/OG
CoG/stn
stn
CoG/stn
CoG
CoG/stn
CoG/stn
CoG/stn
1554.6972
RFamide
948.5414
965.5428
1005.5741
1019.5897
1022.5643
1073.5527
1104.6061
1119.6455
1124.6323
1147.6483
1158.6167
1238.6640
1271.6394
1287.6844
1288.6797
1314.7753
CabTRP
934.4927
1026.5189
SIFa
1381.7375
CPRP
1033.5207
1478.7485
Pyrokinin
878.5247
1037.5527
RYamide
976.4635
1030.4508
YRamide
844.4788
a
NFDEIDRTGFGFH
+
-
-
CoG/stn
PGVNFLRFa
NRNFLRFa
GRPNFLRFa
APRNFLRFa
GNRNFLRFa
TNYGGFLRFa
GAHKNYLRFa
SMPTLRLRFa
GLSRNYLRFa
APQRNFLRFa
YGNRSFLRFa
SQPSKNYLRFa
pQDLDHVFLRFa
GYSVGLNYLRFa
QDLDHVFLRFa
DARTAPLRLRFa
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
-
+
+
+
+
+
+
+
APSGFLGMRa
YPSGFLGMRa
+
+
-
+
+
CoG/stn
CoG
GYRKPPFNGSIFa
+
+
+
CoG/stn
RSAEGLGRMG
TPLGDLSGSLGHPVE
+
+
+
-
stn
LYFAPRLamide
SGGFAFSPRLa
+
-
+
-
CoG
stn
SGFYANRYa
pEGFYSQRYa
-
-
-
CoG
CoG
HIGSLYRa
+
-
+
CoG
CoG/stn
stn
stn
CoG/stn
CoG/stn
CoG/stn
CoG/stn
CoG/stn
CoG/stn
CoG
CoG
CoG
CoG/stn
The neuropeptides identified from other locations of STNS were conducted on MALDI-TOF/TOF.
3
Supplemental Figure S1. Representative tandem mass spectra highlight MS/MS
sequencing of two neuropeptides obtained directly from the C. sapidus STG. The amino acid
sequence of each peptide is given above the spectrum. (a) MS/MS fragmentation of orcokinin
NFDEIDRSGFGFA (m/z 1474.66). (b) MS/MS fragmentation of YRamide HIGSLYRa (m/z
844.48). The presence of b and y ions is indicated by horizontal lines above (y ions) or below (b
ions) the corresponding amino acid residues in the peptide sequence. The fragment ions resulting
from a lipid with close mass to the YRamide are labeled in blue.
4
Supplemental Figure S2. Collision-induced dissociation spectra of neuropeptides from the
STG extract of C. sapidus on nanoLC-ESI-Q-TOF instrument. (a) MS/MS fragmentation of
CabTRP 1a APSGFLGMRa (m/z 934.49). (b) MS/MS fragmentation of SIFamide
GYRKPPFNGSIFa (m/z 1381.74). The presence of b and y ions is indicated by horizontal lines
above (b ions) or below (y ions) the corresponding amino acid residues in the peptide sequence.
5
Supplemental Figure S3. MALDI TOF/TOF imaging of the C. sapidus STNS. Distribution
of three neuropeptides are shown, including (a) YRamide HIGSLYRa (m/z 844.48) and two
orcokinins: (b) NFDEIDRSSFa (m/z 1228.56) and (c) NFDEIDRSSFGFV (m/z 1532.70); and the
MS image of a potassiated lipid PC(36:1) at m/z 826.57 is displayed in (d).
6