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Volume 13
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Article

Screening and Characterization of Sialic Acid-Binding Variable Lymphocyte Receptors from Hagfish

by
Mark Rickard N. Angelia
1,2,
Abigail Joy D. Rodelas-Angelia
1,
Cheolung Yang
1,
Sojeong Park
1,
Seung pyo Jeong
1,
Hyeok Jang
1,
Dennis Berbulla Bela-ong
1,
Hobin Jang
3,
Kim D. Thompson
4 and
Taesung Jung
1,*
1
Laboratory of Aquatic Animal Diseases, Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, 501-201, 501, Jinju-Daero, Jinju-si 52828, Gyeongsangnam-do, Republic of Korea
2
Institute of Chemistry, College of Arts and Sciences, University of the Philippines Los Baños, College, Laguna 4031, Philippines
3
Center for Study of Emerging and Re-Emerging Viruses, Korea Virus Research Institute, Institute for Basic Science, Daejeon 34126, Republic of Korea
4
Moredun Research Institute, Pentlands Science Park, Bush Loan, Midlothian EH26 0PZ, UK
*
Author to whom correspondence should be addressed.
BioTech 2024, 13(4), 46; https://doi.org/10.3390/biotech13040046
Submission received: 2 September 2024 / Revised: 29 October 2024 / Accepted: 8 November 2024 / Published: 12 November 2024
(This article belongs to the Section Agricultural and Food Biotechnology)
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Abstract

Sialic acid is a diverse group of monosaccharides often found on the termini of N- and O-linked glycans as well as being components of glycoconjugates. Hypersialylation has been associated with the progression of chronic inflammation-mediated diseases such as cardiovascular disease and cancer. Given its role in infection and disease-related processes, sialic acid is a promising target for therapeutic approaches that utilize carbohydrate-binding molecules. In this study, we screened for sialic acid-recognizing variable lymphocyte receptors (VLRBs) or ccombodies from inshore hagfish (Eptatretus burgeri) using a synthetic Neu5Ac-glycoconjugate as an antigen in immunoassay. Resulting ccombodies, 2D8, 5G11, 4A1, and 5F8 were further characterized in terms of their binding activity and specificity. A competitive ELISA using free haptens showed strong inhibition using either N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc). The half-maximal inhibitory concentrations (IC50) for Neu5Ac ranged from 7.02 to 17.06 mM, with candidates 4A1 and 5G11 requiring the least and highest amounts, respectively. IC50 values for Neu5Gc ranged from 8.12 to 13.91 mM, for 4A1 and 5G11, respectively. Candidate ccombodies also detected naturally occurring sialic acid from known sialoglycoproteins using a dot blot assay. Neu5Gc-5G11 and Neu5Ac-2D8 yielded the strongest and weakest docking interactions with affinity values of −5.9 kcal/mol and −4.9 kcal/mol, respectively. Hydrogen bonding and hydrophobic interactions were predicted to be the predominant noncovalent forces observed between the ccombodies and sialic acid. This study demonstrates that glycan-binding VLRBs from hagfish hold promise in augmenting the glycobiologists’ toolkit in investigating the roles of glycans in human and animal health and disease.
Keywords: variable lymphocyte receptors; hagfish VLRB; sialic acid binding; glycan recognition; ccombodies variable lymphocyte receptors; hagfish VLRB; sialic acid binding; glycan recognition; ccombodies

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MDPI and ACS Style

Angelia, M.R.N.; Rodelas-Angelia, A.J.D.; Yang, C.; Park, S.; Jeong, S.p.; Jang, H.; Bela-ong, D.B.; Jang, H.; Thompson, K.D.; Jung, T. Screening and Characterization of Sialic Acid-Binding Variable Lymphocyte Receptors from Hagfish. BioTech 2024, 13, 46. https://doi.org/10.3390/biotech13040046

AMA Style

Angelia MRN, Rodelas-Angelia AJD, Yang C, Park S, Jeong Sp, Jang H, Bela-ong DB, Jang H, Thompson KD, Jung T. Screening and Characterization of Sialic Acid-Binding Variable Lymphocyte Receptors from Hagfish. BioTech. 2024; 13(4):46. https://doi.org/10.3390/biotech13040046

Chicago/Turabian Style

Angelia, Mark Rickard N., Abigail Joy D. Rodelas-Angelia, Cheolung Yang, Sojeong Park, Seung pyo Jeong, Hyeok Jang, Dennis Berbulla Bela-ong, Hobin Jang, Kim D. Thompson, and Taesung Jung. 2024. "Screening and Characterization of Sialic Acid-Binding Variable Lymphocyte Receptors from Hagfish" BioTech 13, no. 4: 46. https://doi.org/10.3390/biotech13040046

APA Style

Angelia, M. R. N., Rodelas-Angelia, A. J. D., Yang, C., Park, S., Jeong, S. p., Jang, H., Bela-ong, D. B., Jang, H., Thompson, K. D., & Jung, T. (2024). Screening and Characterization of Sialic Acid-Binding Variable Lymphocyte Receptors from Hagfish. BioTech, 13(4), 46. https://doi.org/10.3390/biotech13040046

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