A biotechnological tool for glycoprotein desialylation based on immobilized neuraminidase from Clostridium perfringens
Background: Sialic acids are widely distributed in nature and have biological relevance owing to their varied structural and functional roles. Immobilized neuraminidase can selectively remove terminal N-acetyl neuraminic acid from glycoproteins without altering the protein backbone while it can be e...
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| Format: | Article |
| Language: | English |
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Elsevier
2021-07-01
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| Series: | Biochemistry and Biophysics Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405580821000340 |
| _version_ | 1798034463580487680 |
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| author | Lucía Bidondo Mercedes Landeira Florencia Festari Teresa Freire Cecilia Giacomini |
| author_facet | Lucía Bidondo Mercedes Landeira Florencia Festari Teresa Freire Cecilia Giacomini |
| author_sort | Lucía Bidondo |
| collection | DOAJ |
| description | Background: Sialic acids are widely distributed in nature and have biological relevance owing to their varied structural and functional roles. Immobilized neuraminidase can selectively remove terminal N-acetyl neuraminic acid from glycoproteins without altering the protein backbone while it can be easily removed from the reaction mixture avoiding sample contamination. This enables the evaluation of changes in glycoprotein performance upon desialylation. Methods: Neuraminidase was immobilized onto agarose activated with cyanate ester groups and further used for desialylation of model glycoproteins, a lysate from tumour cells and tumour cells. Desialylation process was analysed by lectin binding assay, determination of sialyl-Tn or flow cytometry. Results: Clostridium perfringens neuraminidase was immobilized with 91 % yield and expressed activity yield was of 41%. It was effective in the desialylation of bovine fetal serum fetuin, bovine lactoferrin and ovine submaxilar mucin. A decrease in sialic-specific SNA lectin recognition of 83% and 53 % was observed for fetuin and lactoferrin with a concomitant increase in galactose specific ECA and PNA lectin recognition. Likewise, a decrease in the recognition of a specific antibody (82%) upon mucin desialylation was observed. Moreover, desialylation of a protein lysate from the sialic acid-rich cell line TA3/Ha was also possible leading to a decrease in 47 % in SNA recognition. Immobilized neuraminidase kept 100% of its initial activity upon five desialylation cycles. Conclusions: Immobilized neuraminidase is an interesting as well as a robust biotechnological tool for enzymatic desialylation purposes. General significance: Immobilized neuraminidase would contribute to understand the role of sialic acid in biological processes. |
| first_indexed | 2024-04-11T20:44:36Z |
| format | Article |
| id | doaj.art-a89c4701a3bd4f4e86152cae625653a0 |
| institution | Directory Open Access Journal |
| issn | 2405-5808 |
| language | English |
| last_indexed | 2024-04-11T20:44:36Z |
| publishDate | 2021-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biochemistry and Biophysics Reports |
| spelling | doaj.art-a89c4701a3bd4f4e86152cae625653a02022-12-22T04:04:05ZengElsevierBiochemistry and Biophysics Reports2405-58082021-07-0126100940A biotechnological tool for glycoprotein desialylation based on immobilized neuraminidase from Clostridium perfringensLucía Bidondo0Mercedes Landeira1Florencia Festari2Teresa Freire3Cecilia Giacomini4Laboratorio de Bioquímica, Departamento de Biociencias, Facultad de Química, UdelaR, Gral. Flores, 2124, Montevideo, UruguayLaboratorio de Inmunomodulación y desarrollo de Vacunas, Departamento de Inmunobiología, Facultad de Medicina, UdelaR, Gral Flores, 2125, Montevideo, UruguayLaboratorio de Inmunomodulación y desarrollo de Vacunas, Departamento de Inmunobiología, Facultad de Medicina, UdelaR, Gral Flores, 2125, Montevideo, UruguayLaboratorio de Inmunomodulación y desarrollo de Vacunas, Departamento de Inmunobiología, Facultad de Medicina, UdelaR, Gral Flores, 2125, Montevideo, UruguayLaboratorio de Bioquímica, Departamento de Biociencias, Facultad de Química, UdelaR, Gral. Flores, 2124, Montevideo, Uruguay; Corresponding author.Background: Sialic acids are widely distributed in nature and have biological relevance owing to their varied structural and functional roles. Immobilized neuraminidase can selectively remove terminal N-acetyl neuraminic acid from glycoproteins without altering the protein backbone while it can be easily removed from the reaction mixture avoiding sample contamination. This enables the evaluation of changes in glycoprotein performance upon desialylation. Methods: Neuraminidase was immobilized onto agarose activated with cyanate ester groups and further used for desialylation of model glycoproteins, a lysate from tumour cells and tumour cells. Desialylation process was analysed by lectin binding assay, determination of sialyl-Tn or flow cytometry. Results: Clostridium perfringens neuraminidase was immobilized with 91 % yield and expressed activity yield was of 41%. It was effective in the desialylation of bovine fetal serum fetuin, bovine lactoferrin and ovine submaxilar mucin. A decrease in sialic-specific SNA lectin recognition of 83% and 53 % was observed for fetuin and lactoferrin with a concomitant increase in galactose specific ECA and PNA lectin recognition. Likewise, a decrease in the recognition of a specific antibody (82%) upon mucin desialylation was observed. Moreover, desialylation of a protein lysate from the sialic acid-rich cell line TA3/Ha was also possible leading to a decrease in 47 % in SNA recognition. Immobilized neuraminidase kept 100% of its initial activity upon five desialylation cycles. Conclusions: Immobilized neuraminidase is an interesting as well as a robust biotechnological tool for enzymatic desialylation purposes. General significance: Immobilized neuraminidase would contribute to understand the role of sialic acid in biological processes.http://www.sciencedirect.com/science/article/pii/S2405580821000340NeuraminidaseSialic acidImmobilizationGlycomic analysisEnzymatic desialylation |
| spellingShingle | Lucía Bidondo Mercedes Landeira Florencia Festari Teresa Freire Cecilia Giacomini A biotechnological tool for glycoprotein desialylation based on immobilized neuraminidase from Clostridium perfringens Biochemistry and Biophysics Reports Neuraminidase Sialic acid Immobilization Glycomic analysis Enzymatic desialylation |
| title | A biotechnological tool for glycoprotein desialylation based on immobilized neuraminidase from Clostridium perfringens |
| title_full | A biotechnological tool for glycoprotein desialylation based on immobilized neuraminidase from Clostridium perfringens |
| title_fullStr | A biotechnological tool for glycoprotein desialylation based on immobilized neuraminidase from Clostridium perfringens |
| title_full_unstemmed | A biotechnological tool for glycoprotein desialylation based on immobilized neuraminidase from Clostridium perfringens |
| title_short | A biotechnological tool for glycoprotein desialylation based on immobilized neuraminidase from Clostridium perfringens |
| title_sort | biotechnological tool for glycoprotein desialylation based on immobilized neuraminidase from clostridium perfringens |
| topic | Neuraminidase Sialic acid Immobilization Glycomic analysis Enzymatic desialylation |
| url | http://www.sciencedirect.com/science/article/pii/S2405580821000340 |
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