Identification and functional analysis of a novel L-type lectin (NdLTL1) from Neocaridina denticulata sinensis.

This study investigates an L-type lectin, NdLTL1, derived from Neocaridina denticulata sinensis, emphasizing its role in immune defense through carbohydrate binding and bacterial agglutination. Bioinformatics analysis identified 179 lectin sequences, leading to subsequent investigations into the structure and function of NdLTL1. The open reading frame (ORF) of NdLTL1 spans 966 bp and encodes a protein consisting of 321 amino acids (36.25 kDa), which features a signal peptide, a transmembrane domain and Lectin_leg-like domain. Three-dimensional modeling revealed three antiparallel β-sheets characteristic of Lectin_leg-like domain, confirming evolutionary links with proteins such as VIP36. Protein-carbohydrate and protein-protein interaction studies showed that NdLTL1 binds to both carbohydrates like N-acetylglucosamine, peptidoglycan, lipopolysaccharides (LPS), and mannose, as well as sorting proteins (COPI/COPII). Gene expression analyses indicated that NdLTL1 exhibits the highest expression levels in cardiac tissues and significant upregulation in gills following exposure to Vibrio parahaemolyticus. Recombinant NdLTL1 expressed in Escherichia coli was shown to bind multiple bacterial strains and exhibit calcium-dependent agglutination properties. Enzyme-linked immunosorbent assay (ELISA) confirmed concentration-dependent carbohydrate binding, particularly rapid for LPS. In vitro experiments suggested that recombinant NdLTL1 may promote bacterial growth under nutrient-limited conditions while potentially triggering immune defenses indirectly.