Molecular characterization and antibacterial ability of galectin-3 and galectin-9 in Onychostoma macrolepis

Hongzhou Xu; Haixia Liu; Chengrong Liu; Xinyan Shangguan; Xu Cheng; Ruifang Zhang; Yitong Lu; Ping Li; Yingjie Cai

doi:10.1016/j.dci.2021.104333

Molecular characterization and antibacterial ability of galectin-3 and galectin-9 in Onychostoma macrolepis

Dev Comp Immunol. 2022 Mar:128:104333. doi: 10.1016/j.dci.2021.104333. Epub 2021 Dec 13.

Authors

Hongzhou Xu¹, Haixia Liu², Chengrong Liu¹, Xinyan Shangguan¹, Xu Cheng¹, Ruifang Zhang¹, Yitong Lu¹, Ping Li³, Yingjie Cai¹

Affiliations

¹ College of Animal Science and Technology, Northwest A&F University, Xinong Road 22nd, Yangling, Shaanxi, 712100, PR China.
² College of Animal Science and Technology, Northwest A&F University, Xinong Road 22nd, Yangling, Shaanxi, 712100, PR China. Electronic address: liuhaixia209@nwafu.edu.cn.
³ Centre for Research on Environmental Ecology and Fish Nutrition (CREEFN) of the Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, 201306, PR China.

PMID: 34914929
DOI: 10.1016/j.dci.2021.104333

Abstract

Galectins belong to the β-galactoside binding protein family, which have conserved carbohydrate-recognition domains (CRDs) and participate in innate and acquired immunity in animals. In this study, two galectin genes were cloned from Onychostoma macrolepis, OmGal-3 (galectin-3) and OmGal-9 (galectin-9). The open reading frames (ORFs) of OmGal-3 and OmGal-9 contain 732 and 978 base pairs, encoding 243 and 325 amino acids, respectively. OmGal-3 contains a C-terminal CRD, but OmGal-9 contains an N-terminal CRD and a C-terminal CRD. Two galectins were expressed at varying levels in all tissues examined, with the liver showing the highest expression. The relative gene expression levels of OmGal-3 and OmGal-9 following Aeromonas hydrophila infection were significantly up-regulated in the liver and spleen, and OmGal-9 had a greater increase than OmGal-3. The recombinant OmGal-3 and OmGal-9 proteins (rOmGal-3 and rOmGal-9) were authenticated and verified by SDS-PAGE and western blotting. ROmGal-3 and rOmGal-9 agglutinated all tested bacteria, including 3 g-positive bacteria (Aeromonas hydrophila, Escherichia coli, and Vibrio parahaemolyticus) and 3 g-negative bacteria (Streptococcus agalactiae, Staphylococcus aureus, and Bacillus cereus) in vivo without Ca²⁺. ROmGal-3 showed strong binding both to gram-positive and gram-negative bacteria and OmGal-9 had a stronger binding activity against gram-positive bacteria. Furthermore, rOmGal-3 and rOmGal-9 exhibited dose-dependent binding capability to two classic pathogens associated molecular pattern (LPS and PGN) and two sugars (d-lactose and d-galactose), and rOmGal-3 has better binding activity at lower concentrations in LPS and PGN than rOmGal-3. The integrated analyses indicate that the two galectins probably play an important role in innate immune defense by binding to bacterial cells via the CRD domain against pathogen infection.

Keywords: Agglutinating activity; Bacteria infection; Galectin-3; Galectin-9; Onychostoma macrolepis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Anti-Bacterial Agents* / pharmacology
Cyprinidae* / genetics
Cyprinidae* / physiology
Fish Proteins* / genetics
Fish Proteins* / pharmacology
Galectin 3* / genetics
Galectin 3* / pharmacology
Gene Expression Regulation
Gram-Negative Bacteria / drug effects
Gram-Positive Bacteria / drug effects
Immunity, Innate / genetics
Phylogeny
Sequence Alignment

Substances

Anti-Bacterial Agents
Fish Proteins
Galectin 3