P82 Peptide fraction identification by SE-HPLC and LC-MS/MS analysis of the body mucus from Portugal coastal fish Halobatrachus didactylus

M. Cunha; E. R. Coscueta; M. E. Brassesco; F. Almada; D. Gonçalves; M. Manuela Pintado

P82 Peptide fraction identification by SE-HPLC and LC-MS/MS analysis of the body mucus from Portugal coastal fish Halobatrachus didactylus

M. Cunha^*, E. R. Coscueta, M. E. Brassesco, F. Almada, D. Gonçalves, M. Manuela Pintado

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › peer-review

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Abstract

The mucus covers the fish's body, working as a protective barrier. Besides physical protection, mucus provides molecules that protect the fish from pathogens damaging 1,2. These include antimicrobial peptides secreted in the mucus, which play an essential role in defense against microbial pathogens since these belong to the innate immune system2,3. In this study, two adult Halobatrachus didactylus individuals were captured from the wild in Sesimbra. Then, mucus collection was performed by scraping the dorsal-lateral body of the fish with a sponge. Our objective was the identification of new peptides with bioactive potential in mucus samples by chromatography analysis. Size exclusion highperformance liquid chromatography (SE-HPLC) analysis performed on mucus samples from the two individuals revealed a similar profile with an intense highlight peak which resulted in a distribution of about 775 Dalton. With interest in that peak, the two mucus samples were pooled for fractionation by SEC. The resulting fraction was analyzed by liquid chromatography-tandem mass spectrometry (LCMS/MS) to identify the most probable peptide sequences. Identification from databases did not provide reliable results, indicating a lack of information on the matrix analyzed. We resorted to de novo sequencing with good results using PEAKS Studio software. Five identified peptides were selected according to their bioactivities predicted in silico. Furthermore, the five identified peptides were synthesized, and the molecular size was validated by SE-HPLC analysis. Overall, this chromatographic approach enabled the identification of promising peptides, which bioactivities will be evaluated in vitro in future work.

Original language	English
Title of host publication	Livro de resumos do 12º Encontro Nacional de Cromatografia & XIV WARPA
Editors	Sílvia M. Rocha
Place of Publication	Lisboa
Publisher	Sociedade Portuguesa de Química
Pages	208-208
Number of pages	1
ISBN (Electronic)	9789898124371
Publication status	Published - 6 Dec 2022
Event	12º Encontro Nacional de Cromatografia - Universidade de Aveiro, Aveiro, Portugal Duration: 6 Dec 2022 → 8 Dec 2022 https://12enc.events.chemistry.pt/

Conference

Conference	12º Encontro Nacional de Cromatografia
Abbreviated title	12ENC
Country/Territory	Portugal
City	Aveiro
Period	6/12/22 → 8/12/22
Internet address	https://12enc.events.chemistry.pt/

Keywords

Fish
Mucus
Halobatrachus didactylus
SE-HPLC
LC-MS/MS
Peptides

Access to Document

60163147Final published version, 605 KBLicence: Unspecified

http://hdl.handle.net/10400.14/39820Licence: Unspecified

1 Participating in a conference, workshop, ...

12º Encontro Nacional de Cromatografia
Marta Fernandez da Cunha (Participant)
6 Dec 2022 → 8 Dec 2022
Activity: Participating in or organising an event › Participating in a conference, workshop, ...

1 Active

CBQF Multianual Funding: Centre for Biotechnology and Fine Chemistry
Cortez, J., Pintado, M. M., Morais, A. M. M. B., Ribeiro, A. B., Ferreira, A. C., Castro, P., Villamor, A. L., Oliveira, A. L., Oliveira, A. L. S., Amaro, A. L., Paulo, A., Gomes, A. M., Marques, A., Carvalho, A. P., Monforte, A. R., Madureira, A. R., Oliveira, A. S., Ferreira, A. S., Rangel, A. O. S. S., Oliveira, C., Cálix, C., Pereira, C. F., Santos, C. S. D., Amorim, C. L., Oliveira, C., Manaia, C., Silva, C. L. M., Monteiro, C. M., Machado, D., Campos, D., Oliveira, D., Valente, D., Cardoso, E., Costa, E. M. A. D., Pinto, E., Alexandre, E., Carsanba, E., Coscueta, E., Raposo, M. F. D. J., Tavaria, F., Voss, G. B., Moreira, H., Campos, I., Tomada, I., Moreira, I. S., Vaz-Moreira, I., Fundo, J. F., Barbosa, J., Pereira, J. O., Durão, J. R. D. O., Costa, J. R., Fernandes, J. C., Ferreira, J., Silva, J. A., Couto, J. A., Oliveira, L., Pimentel, L., Rodríguez-Alcalá, L. M., Hogg, C., Poças, M. D. F., Miller, F. A., Brassesco, M. E., Mota, I., Ramos, I. N., Vasconcelos, I., Monteiro, M. J., Amorim, M., Roriz, M., Silva, M., Correia, M., Silva, M. N. D., Alves, M., Vasconcelos, M., Ramos, O. L., Costa, P., Gibbs, P., Teixeira, P., Ramos, P., Rodrigues, P. M., Carvalho, P., Mesquita, R., Barros, R., Morais, R. M. S. C., Magalhães, R. M. B. D. S., Borges, S. C. F., Correia, S., Silva, S. B. D., Silva, S., Moreira, S. A. M., Pedrosa, S., Pereira, S., Vidigal, S., Pedro, T., Ribeiro, T., Deuchande, T., Brandão, T. R. S., Resende, T., Hogg, T., Ferreira, V., Viseras, A. D., Soares, A. M. S., Oliveira, A. S., Uribe, B., Pais-Vieira, C., Ferreira, C., Sousa, C. C. S., Kumla, D., Carsanba, E., Lopes, G. L. L., Ribeiro, I. M. M. V. P., Cruz, I. B. D., Barbosa, J. C., Oliveira, L., Mendes, M. A. R., Boas, A. V., Cassoni, A. C., Lopes, A., Pintado, A. I. E., Sousa, A. P., Faustino, A., Salsinha, A. S., Couto, A. T., Macieira, A., Horta, B. M., Silva, C. M. E., Rodrigues, C. M., Bernardes, B. G., Silva, B. I. Q. L. D., Costa, C., Costa, C. M., Serafim, C., Santos, D., Antunes, F., Vieira, E., Ferreira, H., Moreira, I. A., Vieira, I. R. D. F., Rocha, J., Henggeler , J., Miranda, J., Oliveira, M., Lopes, M. T. B. V., Silva, N., Carvalho, N., Costa, P., Nova, P., Sousa, P., Silva, P., Gómez-García, R., Freixo, R., Marçal, S., Cunha, S. A., Sousa, S., Pereira, S. A., Ghalamara, S., Vilela, T., Lopes, A., Lopes, A. I. R., Linhares, A. L. S., Boas, A. M. M. V., Teixeira, A. M. R., Mendes, A. R. M., Oliveira, A. S., Sousa, A. S. D. S., Sousa, A. S. D. S., Martins, A. S. P. D. P., Veiga, A., Bernardes, B. G., Silva, B. I. Q. L. D., Fernandes, A. S., Miranda, C., Dias, C. L. A., Neto, C. C., Castro, C. M. O., Oliveira, D. F., Rodrigues, D., Magalhães, D. D. S., Almeida, D., Castro, D., Costa, E. C. L. D., Darú, F. A., Bastos, F., Teixeira, F. D. S., Ferreira, F., Fortunato, G., Araújo-Rodrigues, H., Osorio Pérez, J. R., Silveira, J. F., Silva, J. A., Soares, J. C., Carvalho, L. C., Castro, L. M. G., Machado, M., Carvalho, M. J., Nazareth, M., Veiga, M., Coelho, M., Carvalho, M. I. P. D., Mendes, M., Ramos, M. A. M., Silva, T., Resende, T., Fontes, A. L., Neves, D. I. S. C. D., Cachetas, D. M., Silva, G. A. D. R. & Morais, R. M. S. C.
1/01/20 → 31/12/23
Project: Research

1 Poster

Peptide fraction identification by SE-HPLC and LC-MS/MS analysis of the body mucus from Portugal coastal fish Halobatrachus didactylus
Cunha, M., Coscueta, E. R., Brassesco, M. E., Almada, F., Gonçalves, D. & Pintado, M., Dec 2022.
Research output: Contribution to conference › Poster

Open Access
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Cite this

Cunha, M., Coscueta, E. R., Brassesco, M. E., Almada, F., Gonçalves, D., & Pintado, M. M. (2022). P82 Peptide fraction identification by SE-HPLC and LC-MS/MS analysis of the body mucus from Portugal coastal fish Halobatrachus didactylus. In S. M. Rocha (Ed.), Livro de resumos do 12º Encontro Nacional de Cromatografia & XIV WARPA (pp. 208-208). Sociedade Portuguesa de Química. http://hdl.handle.net/10400.14/39820

@inproceedings{35af46ec31df4e1091c348c955e6ba0f,

title = "P82 Peptide fraction identification by SE-HPLC and LC-MS/MS analysis of the body mucus from Portugal coastal fish Halobatrachus didactylus",

abstract = "The mucus covers the fish's body, working as a protective barrier. Besides physical protection, mucus provides molecules that protect the fish from pathogens damaging 1,2. These include antimicrobial peptides secreted in the mucus, which play an essential role in defense against microbial pathogens since these belong to the innate immune system2,3. In this study, two adult Halobatrachus didactylus individuals were captured from the wild in Sesimbra. Then, mucus collection was performed by scraping the dorsal-lateral body of the fish with a sponge. Our objective was the identification of new peptides with bioactive potential in mucus samples by chromatography analysis. Size exclusion highperformance liquid chromatography (SE-HPLC) analysis performed on mucus samples from the two individuals revealed a similar profile with an intense highlight peak which resulted in a distribution of about 775 Dalton. With interest in that peak, the two mucus samples were pooled for fractionation by SEC. The resulting fraction was analyzed by liquid chromatography-tandem mass spectrometry (LCMS/MS) to identify the most probable peptide sequences. Identification from databases did not provide reliable results, indicating a lack of information on the matrix analyzed. We resorted to de novo sequencing with good results using PEAKS Studio software. Five identified peptides were selected according to their bioactivities predicted in silico. Furthermore, the five identified peptides were synthesized, and the molecular size was validated by SE-HPLC analysis. Overall, this chromatographic approach enabled the identification of promising peptides, which bioactivities will be evaluated in vitro in future work.",

keywords = "Fish, Mucus, Halobatrachus didactylus, SE-HPLC, LC-MS/MS, Peptides",

author = "M. Cunha and Coscueta, {E. R.} and Brassesco, {M. E.} and F. Almada and D. Gon{\c c}alves and Pintado, {M. Manuela}",

year = "2022",

month = dec,

day = "6",

language = "English",

pages = "208--208",

editor = "Rocha, {S{\'i}lvia M.}",

booktitle = "Livro de resumos do 12º Encontro Nacional de Cromatografia & XIV WARPA",

publisher = "Sociedade Portuguesa de Qu{\'i}mica",

note = "12º Encontro Nacional de Cromatografia, 12ENC ; Conference date: 06-12-2022 Through 08-12-2022",

url = "https://12enc.events.chemistry.pt/",

}

Cunha, M , Coscueta, ER , Brassesco, ME, Almada, F, Gonçalves, D & Pintado, MM 2022, P82 Peptide fraction identification by SE-HPLC and LC-MS/MS analysis of the body mucus from Portugal coastal fish Halobatrachus didactylus. in SM Rocha (ed.), Livro de resumos do 12º Encontro Nacional de Cromatografia & XIV WARPA. Sociedade Portuguesa de Química, Lisboa, pp. 208-208, 12º Encontro Nacional de Cromatografia, Aveiro, Portugal, 6/12/22. <http://hdl.handle.net/10400.14/39820>

P82 Peptide fraction identification by SE-HPLC and LC-MS/MS analysis of the body mucus from Portugal coastal fish Halobatrachus didactylus. / Cunha, M.; Coscueta, E. R.; Brassesco, M. E. et al.

Livro de resumos do 12º Encontro Nacional de Cromatografia & XIV WARPA. ed. / Sílvia M. Rocha. Lisboa : Sociedade Portuguesa de Química, 2022. p. 208-208.

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › peer-review

TY - GEN

T1 - P82 Peptide fraction identification by SE-HPLC and LC-MS/MS analysis of the body mucus from Portugal coastal fish Halobatrachus didactylus

AU - Cunha, M.

AU - Coscueta, E. R.

AU - Brassesco, M. E.

AU - Almada, F.

AU - Gonçalves, D.

AU - Pintado, M. Manuela

PY - 2022/12/6

Y1 - 2022/12/6

N2 - The mucus covers the fish's body, working as a protective barrier. Besides physical protection, mucus provides molecules that protect the fish from pathogens damaging 1,2. These include antimicrobial peptides secreted in the mucus, which play an essential role in defense against microbial pathogens since these belong to the innate immune system2,3. In this study, two adult Halobatrachus didactylus individuals were captured from the wild in Sesimbra. Then, mucus collection was performed by scraping the dorsal-lateral body of the fish with a sponge. Our objective was the identification of new peptides with bioactive potential in mucus samples by chromatography analysis. Size exclusion highperformance liquid chromatography (SE-HPLC) analysis performed on mucus samples from the two individuals revealed a similar profile with an intense highlight peak which resulted in a distribution of about 775 Dalton. With interest in that peak, the two mucus samples were pooled for fractionation by SEC. The resulting fraction was analyzed by liquid chromatography-tandem mass spectrometry (LCMS/MS) to identify the most probable peptide sequences. Identification from databases did not provide reliable results, indicating a lack of information on the matrix analyzed. We resorted to de novo sequencing with good results using PEAKS Studio software. Five identified peptides were selected according to their bioactivities predicted in silico. Furthermore, the five identified peptides were synthesized, and the molecular size was validated by SE-HPLC analysis. Overall, this chromatographic approach enabled the identification of promising peptides, which bioactivities will be evaluated in vitro in future work.

AB - The mucus covers the fish's body, working as a protective barrier. Besides physical protection, mucus provides molecules that protect the fish from pathogens damaging 1,2. These include antimicrobial peptides secreted in the mucus, which play an essential role in defense against microbial pathogens since these belong to the innate immune system2,3. In this study, two adult Halobatrachus didactylus individuals were captured from the wild in Sesimbra. Then, mucus collection was performed by scraping the dorsal-lateral body of the fish with a sponge. Our objective was the identification of new peptides with bioactive potential in mucus samples by chromatography analysis. Size exclusion highperformance liquid chromatography (SE-HPLC) analysis performed on mucus samples from the two individuals revealed a similar profile with an intense highlight peak which resulted in a distribution of about 775 Dalton. With interest in that peak, the two mucus samples were pooled for fractionation by SEC. The resulting fraction was analyzed by liquid chromatography-tandem mass spectrometry (LCMS/MS) to identify the most probable peptide sequences. Identification from databases did not provide reliable results, indicating a lack of information on the matrix analyzed. We resorted to de novo sequencing with good results using PEAKS Studio software. Five identified peptides were selected according to their bioactivities predicted in silico. Furthermore, the five identified peptides were synthesized, and the molecular size was validated by SE-HPLC analysis. Overall, this chromatographic approach enabled the identification of promising peptides, which bioactivities will be evaluated in vitro in future work.

KW - Fish

KW - Mucus

KW - Halobatrachus didactylus

KW - SE-HPLC

KW - LC-MS/MS

KW - Peptides

M3 - Conference contribution

SP - 208

EP - 208

BT - Livro de resumos do 12º Encontro Nacional de Cromatografia & XIV WARPA

A2 - Rocha, Sílvia M.

PB - Sociedade Portuguesa de Química

CY - Lisboa

T2 - 12º Encontro Nacional de Cromatografia

Y2 - 6 December 2022 through 8 December 2022

ER -

P82 Peptide fraction identification by SE-HPLC and LC-MS/MS analysis of the body mucus from Portugal coastal fish Halobatrachus didactylus

Abstract

Conference

Keywords

Access to Document

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Activities

12º Encontro Nacional de Cromatografia

Projects

CBQF Multianual Funding: Centre for Biotechnology and Fine Chemistry

Research output

Peptide fraction identification by SE-HPLC and LC-MS/MS analysis of the body mucus from Portugal coastal fish Halobatrachus didactylus

Cite this