First characterization of fucosidases in spiders

Abstract

l-fucose is a constituent of glycoconjugates in different organisms. Fucosidases catalyze the removal of fucose residues, and have been correlated to different physiological and pathological processes, such as fertilization, cancer, fucosidosis, and digestion in molluscs and ticks. An α-l-fucosidase sequence was identified from the transcriptome and proteome from the midgut diverticula of the synanthropic spider Nephilingis cruentata. In this article, we describe the isolation of this α-l-fucosidase and the characterization of its activity using substrates and inhibitors demonstrating different specificities among fucosidases. The enzyme had a K of 32 and 400 μM for 4-methylumbelliferyl α-l-fucopyranoside and 4-nitrophenyl α-l-fucopyranoside, respectively; and was unable to hydrolyze fucoidan. Nephilingis cruentata α-l-fucosidase was inhibited competitively by fucose and fuconojyrimycin. The fucosidase had two distinct pH optima even in the isolated form, due to oligomerization dependent on pH, as previously described to other fucosidases. Alignment and molecular homology modeling of the protein sequence with other fucosidases indicated that the active sites and catalytic residues were different, including residues involved in acid/base catalysis. Phylogenetic analysis showed, for the first time, gene-duplication events for fucosidases in Arachnida species. All these data reveal that studies on fucosidases in organisms distinct from bacteria, fungi, and humans are important.

Bxy‐fuca encoding α‐L‐fucosidase plays crucial roles in development and reproduction of the pathogenic pinewood nematode, Bursaphelenchus xylophilus

“…In Drosophila , the α‐L‐fucosidase mRNA was detected in the midgut . In addition, digestive α‐L‐fucosidase was found in the midgut or midgut glands in several other species, including Amblyomma sculptum , Nephilingis cruentata , Tityus serrulatus , Pecten maximum , and Biomphalaria glabrata . α‐L‐fucosidase from the plant pathogenic fungus F. graminearum was found actively hydrolyzing the fucose from xyloglucan .…”

Digestive enzymes and sphingomyelinase D in spiders without venom (Uloboridae)

“…Proteomic and biochemical data also identified enzymes involved in the removal of carbohydrates from glycoconjugates, including alpha-L-fucosidase and alpha-mannosidase which are related to the intracellular digestive process in Uloborus sp. The first characterisation of a spider alpha-L-fucosidase was obtained from the midgut of N. cruentata 46 . In insects, the N-glycosylation pathways involve a high mannosylation and two fucosylations (Man3GlcNAc(± α3/6Fuc) GlcNAc) of glycoproteins, forming a structure known as paucimannosidic glycans 47 .…”

Digestive α-L-fucosidase activity in Rhodnius prolixus after blood feeding: effect of secretagogue and nutritional stimuli

“…Conversely, digestive α-L-fucosidases have been identified in molluscs ( Reglero and Cabezas, 1976 ; Endo et al, 1993 ; Berteau et al, 2002 ; Perrella et al, 2015 ; Escobar-Correas et al, 2019 ), ticks ( Moreti et al, 2013 ), spiders ( Perrela et al, 2018b ) and bacteria ( Wu et al, 2021 ; Zhou et al, 2022 ). The knowledge on insect α-L-fucosidases is very scarce, and, despite its initial description and characterization, the role of R. prolixus midgut α-L-fucosidase during blood digestion has not been studied in detail.…”