Accelerated proteomic visualization of individual predatory venoms of Conus purpurascens reveals separately evolved predation-evoked venom cabals
S.W.A. Himaya1, Frank Marí2, Richard J. Lewis1,*
1
IMB Centre for Pain Research, Institute for Molecular Bioscience, The University of Queensland, Queensland 4072, Australia
2
Marine Biochemical Sciences, Chemical Sciences Division, National Institute of Standards and Technology, 331 Fort Johnson Road, Charleston, SC
29412, USA
*
Email:
[email protected]. Phone: +61 7 3346-2984. Fax: +61 7 3346-2101
5
20
0
0
0 0
2467.9
0
5
10
15
20
0
0
Specimen G
10
3056.2
15
20
15
20
Time (min)
Specimen H
PIIIE
100
3988.6
5
Time (min) 4333.6
100
2630.9
10
15
20
0
0
5
Specimen I 2630.9
10
15
Time (min)
0
0
5
10
Time (min)
15
20
0
0
2662.9
5
1348.5
1693.7 3186.4
PVA
2615.9
2614.9
2647.9
1371.5 1458.5
1474.5
9408.1 9392.1
50
9442.1 9424.1
1839.7 1941.7 1839.7 PVA 4976.1 4960.1 1421.5 3186.4
PIVA 4333.6 1474.5 1458.5
20
PIVA
2614.9
3186.4 3055.3 9546.2 9530.2 9497.2
9516.2
3230.4
1348.5
50
PVIIA
4317.6 4377.6
1352.5 2544.06 2675.1
2699.1 1793.7 1777.7
1458.5
1474.5
50 892.3 876.5
PVA 1309.4
3230.4 3299.5 3186.4 3168.4 2731.3
2614.9
6773.2/6772.2 6765.2/6765.7 960.4
4333.6
PIB 2467.9 PIA PIVE
10
Time (min)
5
PVIA
PIVF 2614.9
p6a
3040.2 2997.2
1421.5
PIVA
4333.6 4317.6 4377.6 2630.9 2600.9
2699.1 PVIIA 1777.7
1474.5 1458.5
0
Specimen E
PIIIE
2752.07
Relative intensity (%)
0 0
20
PVIIA
4377.6/4317.6
4317.6 4377.6 2630.9
Time (min)
50
15
4031.9
3005.2
PIIIE
5
100
PVIA p6a/2969.3 3040.2
50
PVIIA
10
PIIIE
100
3598.7
50
Time (min)
Specimen F
2829.08
2871.2/2752.07 1793.7 1777.7/1779.7 PIVA
4333.6
1809.7
1458.5
1474.5
Relative intensity (%)
50
1585.2
PIIIE
2307.8 2527.9
3613.7
5
Time (min) 100
2469.9/2478.9
2483.9
PVIIA
15
2467.9
3020.2
3957.6 3988.6
2407.9
3613.7
2065.7
10
Specimen D
100
PVIA
50
2391.01 2375.01
0 0
2511.03
PVIIA
3002.2 2954.3
2436.9/2435.9
2483.9 4031.9/4046.9
2495.04 2394.9 3374.5
2585.04 2656.01
Specimen C
100
2466.9
3019.3 3024.2 3040.2
2609.07 3411.5
50
PVIA 3002.2 3005.2
5992.5
PVIA 3002.2
2656.06 PIVE
2830.07 4030.9 4046.9
PIVF
Specimen B
100
PVA
Relative intensity (%)
p6a 2969.3
2467.9
Specimen A
100
10
15
20
Time (min)
Figure S1: LC-ESI Triple TOF-MS total ion current chromatogram of the injected predatory venom from nine individual C. purpurascens. LC-MS run on the TripleTOF 5600 System revealed the complexity of the venom.
PVIA
PVIIA
PIVF
PIIIE
PIA
PIVA
PVA
Figure S2: LC-MS/MS interpretations of the known peptides abundantly present in C. purpurascens. Thermo Fisher Scientific Orbitrap Fusion™ Lumos™ Tribrid™ Mass Spectrometer operated in DDA mode was used for data acquisition and MS/MS Data analysis was carried out using the Peaks v8.0 software.
Figure S3: The heatmap matrices of relative expression levels of top 100 (a), 1000 (b) and the total pooled peptidome of 3207 peptides (c). The distinct peptide expression pattern is clearly extends into the minor components of the venom profiles of each specimen.
