Mass spectrometric identification and denovo sequencing of novel conotoxins from vermivorous cone snail (Conus inscriptus), and preliminary screening of its venom for biological activities in vitro and in vivo

Abstract

Venom of Conus inscriptus , a vermivorous cone snail found abundantly in the southern coastal waters was studied to yield conotoxins through proteomic analysis. A total of 37 conotoxins (4 with single disulfide bonds, 20 with two disulfide bonds and 11 three disulfide-bonded peptides) were identified using mass spectrometric analysis. Among them, amino acid sequences of 11 novel conopeptides with one, two and three disulfides belonging to different classes were derived through manual de novo sequencing. Based on the established primary sequence, they were pharmacologically classified into α conotoxins, µ conotoxins and contryphans. Except In1696 all other conopeptides have undergone C-terminal amidation. The natural venom exhibited 50% lethality at 304.82 µg/mL against zebrafish embryo and 130.31 µg/mL against brine shrimp nauplii. The anticonvulsant study of natural venom effectively reduced the locomotor activity against pentylenetetrazole (PTZ) treated zebrafish. This concludes that the venom peptides from Conus inscriptus exhibit potential anticonvulsant function, which leads to the discovery of lead molecules against seizures.

Biological Activity of Cyclic Peptide Extracted from Sphaeranthus amaranthoides Using De Novo Sequencing Strategy by Mass Spectrometry for Cancer

“…CID fragmentation was performed to find the fragmented daughter ions. All the daughter ions obtained were examined for the respective amino acid sequence [ 15 ]. All spectral data were annotated through the mass-spectrometry software Data Analysis 4.1 (Bruker Daltonics, Bremen, Germany).…”

“…The test wells were supplemented with different concentrations (10 ng/mL, 20 ng/mL, 40 ng/mL, 80 ng/mL, and 160 ng/mL) of the peptides, and the toxicity was assessed. The mortality and developmental deformities of the zebrafish larvae were recorded at 24, 48, and 72 hpf [ 15 , 18 ].…”

Unveiling the Diversity and Modifications of Short Peptides in Buthus martensii Scorpion Venom through Liquid Chromatography-High Resolution Mass Spectrometry

“…Simultaneous modification of the N-and C-termini in short peptides is exceptionally rare in natural peptides. Only a limited number of toxins have been identified to possess disulfide bonds spanning approximately 17 amino acids, with modifications at both the C-and N-terminals involving pyroglutamic acid at the C-terminus and amide at the N-terminus [40]. Kuzmin et al synthesized tachyplesin I with N-terminal acetylation and C-terminal amidation modifications, subsequently comparing its activity to that of unmodified tachyplesin I.…”

“…short peptides is exceptionally rare in natural peptides. Only a limited number of toxin have been identified to possess disulfide bonds spanning approximately 17 amino acids with modifications at both the C-and N-terminals involving pyroglutamic acid at th C-terminus and amide at the N-terminus [40]. Kuzmin et al synthesized tachyplesin with N-terminal acetylation and C-terminal amidation modifications, subsequently comparing its activity to that of unmodified tachyplesin I.…”

Biodegradation of Cholesterol by Enterococcus faecium YY01

“…The bacterial genome was sequenced using de novo sequencing technology, and the genome sequence was assembled from scratch using bioinformatics. [30] A PCR was performed using a pair of universal primers: 27F (5 -GAGTTTTGATCCTGTCCAG-3 ) and 1492R (5 -GTTACCTT-GTTACGACTT-3 ) [31]. The 16S rDNA sequence of the strain was compared and analyzed using BLAST through GenBank and the Ribosomal Database Project.…”