Spatial Distribution and Biochemical Characterization of Serine Peptidase Inhibitors in the Venom of the Brazilian Sea Anemone <i>Anthopleura cascaia</i> Using Mass Spectrometry Imaging
Sea anemones are known to produce a diverse array of toxins with different cysteine-rich peptide scaffolds in their venoms. The serine peptidase inhibitors, specifically Kunitz inhibitors, are an important toxin family that is believed to function as defensive peptides, as well as prevent proteolysi...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-08-01
|
| Series: | Marine Drugs |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1660-3397/21/9/481 |
| _version_ | 1797579075811803136 |
|---|---|
| author | Daiane Laise da Silva Rodrigo Valladão Emidio Beraldo-Neto Guilherme Rabelo Coelho Oscar Bento da Silva Neto Hugo Vigerelli Adriana Rios Lopes Brett R. Hamilton Eivind A. B. Undheim Juliana Mozer Sciani Daniel Carvalho Pimenta |
| author_facet | Daiane Laise da Silva Rodrigo Valladão Emidio Beraldo-Neto Guilherme Rabelo Coelho Oscar Bento da Silva Neto Hugo Vigerelli Adriana Rios Lopes Brett R. Hamilton Eivind A. B. Undheim Juliana Mozer Sciani Daniel Carvalho Pimenta |
| author_sort | Daiane Laise da Silva |
| collection | DOAJ |
| description | Sea anemones are known to produce a diverse array of toxins with different cysteine-rich peptide scaffolds in their venoms. The serine peptidase inhibitors, specifically Kunitz inhibitors, are an important toxin family that is believed to function as defensive peptides, as well as prevent proteolysis of other secreted anemone toxins. In this study, we isolated three serine peptidase inhibitors named <i>Anthopleura cascaia</i> peptide inhibitors I, II, and III (ACPI-I, ACPI-II, and ACPI-III) from the venom of the endemic Brazilian sea anemone <i>A. cascaia</i>. The venom was fractionated using RP-HPLC, and the inhibitory activity of these fractions against trypsin was determined and found to range from 59% to 93%. The spatial distribution of the anemone peptides throughout <i>A. cascaia</i> was observed using mass spectrometry imaging. The inhibitory peptides were found to be present in the tentacles, pedal disc, and mesenterial filaments. We suggest that the three inhibitors observed during this study belong to the venom Kunitz toxin family on the basis of their similarity to PI-actitoxin-aeq3a-like and the identification of amino acid residues that correspond to a serine peptidase binding site. Our findings expand our understanding of the diversity of toxins present in sea anemone venom and shed light on their potential role in protecting other venom components from proteolysis. |
| first_indexed | 2024-03-10T22:30:51Z |
| format | Article |
| id | doaj.art-347b74902ff947a2b73b70cd05de9186 |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-03-10T22:30:51Z |
| publishDate | 2023-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-347b74902ff947a2b73b70cd05de91862023-11-19T11:42:11ZengMDPI AGMarine Drugs1660-33972023-08-0121948110.3390/md21090481Spatial Distribution and Biochemical Characterization of Serine Peptidase Inhibitors in the Venom of the Brazilian Sea Anemone <i>Anthopleura cascaia</i> Using Mass Spectrometry ImagingDaiane Laise da Silva0Rodrigo Valladão1Emidio Beraldo-Neto2Guilherme Rabelo Coelho3Oscar Bento da Silva Neto4Hugo Vigerelli5Adriana Rios Lopes6Brett R. Hamilton7Eivind A. B. Undheim8Juliana Mozer Sciani9Daniel Carvalho Pimenta10Programa de Pós-Graduação em Ciências-Toxinologia, Instituto Butantan, Av. Vital Brasil 1500, Butantã, São Paulo 05503-900, BrazilLaboratório de Bioquímica, Instituto Butantan, Av. Vital Brasil 1500, São Paulo 05503-900, BrazilPrograma de Pós-Graduação em Ciências-Toxinologia, Instituto Butantan, Av. Vital Brasil 1500, Butantã, São Paulo 05503-900, BrazilPrograma de Pós-Graduação em Ciências-Toxinologia, Instituto Butantan, Av. Vital Brasil 1500, Butantã, São Paulo 05503-900, BrazilLaboratório de Bioquímica, Instituto Butantan, Av. Vital Brasil 1500, São Paulo 05503-900, BrazilPrograma de Pós-Graduação em Ciências-Toxinologia, Instituto Butantan, Av. Vital Brasil 1500, Butantã, São Paulo 05503-900, BrazilPrograma de Pós-Graduação em Ciências-Toxinologia, Instituto Butantan, Av. Vital Brasil 1500, Butantã, São Paulo 05503-900, BrazilCentre for Microscopy and Microanalysis, University of Queensland, St. Lucia, QLD 4072, AustraliaCentre for Advanced Imaging, University of Queensland, St. Lucia, QLD 4072, AustraliaLaboratório de Farmacologia Molecular e Compostos Bioativos, Universidade São Francisco, Av. São Francisco de Assis, 218, São Paulo 12916-900, BrazilPrograma de Pós-Graduação em Ciências-Toxinologia, Instituto Butantan, Av. Vital Brasil 1500, Butantã, São Paulo 05503-900, BrazilSea anemones are known to produce a diverse array of toxins with different cysteine-rich peptide scaffolds in their venoms. The serine peptidase inhibitors, specifically Kunitz inhibitors, are an important toxin family that is believed to function as defensive peptides, as well as prevent proteolysis of other secreted anemone toxins. In this study, we isolated three serine peptidase inhibitors named <i>Anthopleura cascaia</i> peptide inhibitors I, II, and III (ACPI-I, ACPI-II, and ACPI-III) from the venom of the endemic Brazilian sea anemone <i>A. cascaia</i>. The venom was fractionated using RP-HPLC, and the inhibitory activity of these fractions against trypsin was determined and found to range from 59% to 93%. The spatial distribution of the anemone peptides throughout <i>A. cascaia</i> was observed using mass spectrometry imaging. The inhibitory peptides were found to be present in the tentacles, pedal disc, and mesenterial filaments. We suggest that the three inhibitors observed during this study belong to the venom Kunitz toxin family on the basis of their similarity to PI-actitoxin-aeq3a-like and the identification of amino acid residues that correspond to a serine peptidase binding site. Our findings expand our understanding of the diversity of toxins present in sea anemone venom and shed light on their potential role in protecting other venom components from proteolysis.https://www.mdpi.com/1660-3397/21/9/481serine peptidase inhibitorsvenomsea anemonesmass spectrometry imaging |
| spellingShingle | Daiane Laise da Silva Rodrigo Valladão Emidio Beraldo-Neto Guilherme Rabelo Coelho Oscar Bento da Silva Neto Hugo Vigerelli Adriana Rios Lopes Brett R. Hamilton Eivind A. B. Undheim Juliana Mozer Sciani Daniel Carvalho Pimenta Spatial Distribution and Biochemical Characterization of Serine Peptidase Inhibitors in the Venom of the Brazilian Sea Anemone <i>Anthopleura cascaia</i> Using Mass Spectrometry Imaging Marine Drugs serine peptidase inhibitors venom sea anemones mass spectrometry imaging |
| title | Spatial Distribution and Biochemical Characterization of Serine Peptidase Inhibitors in the Venom of the Brazilian Sea Anemone <i>Anthopleura cascaia</i> Using Mass Spectrometry Imaging |
| title_full | Spatial Distribution and Biochemical Characterization of Serine Peptidase Inhibitors in the Venom of the Brazilian Sea Anemone <i>Anthopleura cascaia</i> Using Mass Spectrometry Imaging |
| title_fullStr | Spatial Distribution and Biochemical Characterization of Serine Peptidase Inhibitors in the Venom of the Brazilian Sea Anemone <i>Anthopleura cascaia</i> Using Mass Spectrometry Imaging |
| title_full_unstemmed | Spatial Distribution and Biochemical Characterization of Serine Peptidase Inhibitors in the Venom of the Brazilian Sea Anemone <i>Anthopleura cascaia</i> Using Mass Spectrometry Imaging |
| title_short | Spatial Distribution and Biochemical Characterization of Serine Peptidase Inhibitors in the Venom of the Brazilian Sea Anemone <i>Anthopleura cascaia</i> Using Mass Spectrometry Imaging |
| title_sort | spatial distribution and biochemical characterization of serine peptidase inhibitors in the venom of the brazilian sea anemone i anthopleura cascaia i using mass spectrometry imaging |
| topic | serine peptidase inhibitors venom sea anemones mass spectrometry imaging |
| url | https://www.mdpi.com/1660-3397/21/9/481 |
| work_keys_str_mv | AT daianelaisedasilva spatialdistributionandbiochemicalcharacterizationofserinepeptidaseinhibitorsinthevenomofthebrazilianseaanemoneianthopleuracascaiaiusingmassspectrometryimaging AT rodrigovalladao spatialdistributionandbiochemicalcharacterizationofserinepeptidaseinhibitorsinthevenomofthebrazilianseaanemoneianthopleuracascaiaiusingmassspectrometryimaging AT emidioberaldoneto spatialdistributionandbiochemicalcharacterizationofserinepeptidaseinhibitorsinthevenomofthebrazilianseaanemoneianthopleuracascaiaiusingmassspectrometryimaging AT guilhermerabelocoelho spatialdistributionandbiochemicalcharacterizationofserinepeptidaseinhibitorsinthevenomofthebrazilianseaanemoneianthopleuracascaiaiusingmassspectrometryimaging AT oscarbentodasilvaneto spatialdistributionandbiochemicalcharacterizationofserinepeptidaseinhibitorsinthevenomofthebrazilianseaanemoneianthopleuracascaiaiusingmassspectrometryimaging AT hugovigerelli spatialdistributionandbiochemicalcharacterizationofserinepeptidaseinhibitorsinthevenomofthebrazilianseaanemoneianthopleuracascaiaiusingmassspectrometryimaging AT adrianarioslopes spatialdistributionandbiochemicalcharacterizationofserinepeptidaseinhibitorsinthevenomofthebrazilianseaanemoneianthopleuracascaiaiusingmassspectrometryimaging AT brettrhamilton spatialdistributionandbiochemicalcharacterizationofserinepeptidaseinhibitorsinthevenomofthebrazilianseaanemoneianthopleuracascaiaiusingmassspectrometryimaging AT eivindabundheim spatialdistributionandbiochemicalcharacterizationofserinepeptidaseinhibitorsinthevenomofthebrazilianseaanemoneianthopleuracascaiaiusingmassspectrometryimaging AT julianamozersciani spatialdistributionandbiochemicalcharacterizationofserinepeptidaseinhibitorsinthevenomofthebrazilianseaanemoneianthopleuracascaiaiusingmassspectrometryimaging AT danielcarvalhopimenta spatialdistributionandbiochemicalcharacterizationofserinepeptidaseinhibitorsinthevenomofthebrazilianseaanemoneianthopleuracascaiaiusingmassspectrometryimaging |