Full length articleCharacterization of a novel O-acetyl sialic acid specific lectin from the hemolymph of the marine crab, Atergatis integerrimus (Lamarck, 1818)
Introduction
Lectins are ubiquitous glycoproteins of non - immune origin that recognize specific carbohydrate structures and agglutinate a variety of animal cells by binding to cell surface glycoproteins and glycolipids [1]. Lectin - carbohydrate interaction represents a ligand - receptor interaction that is universal in all living organisms [2] and such interactions aid in different biological roles like cellular and tissue transport of carbohydrates, glycoproteins and calcium [3], cytolytic and cytotoxic [4] and cell adhesion, migration and apoptosis [5]. They are capable of inducing cell proliferation, cell arrest or apoptosis and have been implicated in organ morphogenesis, tumor cell metastasis, leukocyte trafficking, immune response and inflammation, as well as recognition of extracellular matrix [6]. Lectins isolated from animal tissues were investigated as apoptotic agents, immunomodulatory, antiviral and anticancer drug targets [7]. Among the different classes of lectins isolated from invertebrates, sialic acid specific lectins have gained much importance owing to its function which includes induction of apoptosis, negative regulation of B cell signaling, induction of cytokine secretion [8] and inhibition of bacterial and viral sialidases by altering the immunopotency of sialoconjugates expressed on the microbial surface [9]. Sialic acids are a family of sugars with more than twenty derivatives which differs only in the acyl substitution of the C-5 amino group and O-substituted sialic acids exhibit species and tissue specific distribution [10]. O-acetylation of sialic acids may change with transformation or alteration in the environment of the cell [11] and modified sialic acids such as 9-O-acetyl sialic acid, N-glycolyl neuraminic acid and α-2,6 sialic acids have been detected in human malignant cells [12]. Among arthropods, crustaceans were found to be rich source of sialic acid specific lectins. Lectins have been purified from many brachyuran crabs that are sialic acid specific [13,14]. A 9-O-acetyl and 4-O acetyl sialic acid specific lectin was purified from the hemolymph of marine crab Cancer antennarius which can recognize the modified sugar moieties expressed on tumour cells [15]. A sialic acid specific lectin having a unique specificity for N-glycolyl neuraminic acid has been isolated from marine crab Scylla serrata [16] and multiple agglutinins were detected form S. serrata that agglutinated bacteria [17]. Lectin highly specific to O-acetyl sialic acid and mannose residues expressed on bacterial pathogens was purified from marine crab Erimacrus isenbeckii [18]. A calcium dependent lectin has been isolated from the hemolymph of marine crab Trichopeltarion nobile [19], and Atergatis ocyroe [20] and antimicrobial property of lectin has been demonstrated in marine crabs Portunus pelagicus [21,22]. Lectin that recognizes N-glycolyl neuraminic acid and exhibits antiproliferative property against cancer cells was isolated from the Korean marine crab Philyra pisum and is used as a diagnostic and an anticancer agent [23,24]. The information on isolation and characterization of sialic acid specific lectins from marine crabs is limited. The present investigation reveals purification of novel sialic acid specific lectin from marine crab A. integerrimus that can be used in targeted delivery against microbes and malignant cells.
Section snippets
Collection of hemolymph
The hemolymph from the crab Atergatis integerrimus was collected following the procedure of Mercy and Ravindranath [16]. Marine crabs were collected from the coastal regions of Kanyakumari District and the hemolymph was collected from uninjured, non-autotomised crabs, either by inserting a sterile 1.0 ml syringe with a 22 gauge needle into the arthrodial membrane of the base of third walking leg or by cutting the tip of third walking leg. The hemolymph collected in centrifuge tubes placed on
Purification of Atergatis integerrimus lectin (AiL)
The crude agglutinin Atergatis integerrimus hemolymph was found to have strong agglutinating activity against buffalo erythrocytes. AiL, a lectin from A. integerrimus was purified by bio adsorption using formalinized buffalo erythrocytes and affinity purification (Table 1, Fig. 1, Fig. 2). The specific activity of the affinity purified lectin showed 1218 purification fold and 2.54 × 105 increase in specific activity and formalinized erythrocyte adsorption technique yielded 146 fold purification
Discussion
A sialic acid specific natural lectin - AiL from marine crab Atergatis integerrimus was purified by affinity chromatography using BSM - linked sepharose 4B and bioadsorption using formalinized buffalo erythrocytes. FTIR spectra of the purified lectin of marine crab Atergatis integerrimus exhibited characteristic bands which included both proteins and sugars. The peptide group gave two major bands amide I and amide in the infrared spectrum. The amide I band is associated with the CO stretching
CRediT authorship contribution statement
T. Elayabharathi: Writing - original draft. J. Vinoliya Josephine Mary: Supervision. S. Mary Mettilda Bai: Formal analysis.
References (59)
- et al.
Purification and characterization of a lectin isolated from the Manila clam Ruditapes philippinarum in Korea
Fish Shellfish Immunol.
(2004) - et al.
A cytolytic function for a sialic acid - binding lectin that is a member of the pentraxin family of proteins
J. Biol. Chem.
(1996) - et al.
Lectin- carbohydrate interaction in the immune system
Vet. Immunol. Immunopathol.
(1996) Chemistry, metabolism and biological functions of sialic acids
Adv. Carbohydr. Chem. Biochem.
(1982)- et al.
Purification and characterization of an O-acetyl sialicacid-specific lectin from a marine crab Cancer antennarius
J. Biol. Chem.
(1985) - et al.
Purification, characterization and functional analysis of the immune molecule lectin from the Portunus Pelagicus and their antibiofilm properties
Fish Shellfish Immunol.
(2017) - et al.
Agglutinin from Limulus polyphemus: purification with formalinized horse erythrocytes as the affinity adsorbent
Biochim. Biophys. Acta Protein Struct.
(1975) - et al.
Protein measurement with folin phenol reagent
J. Biol. Chem.
(1951) - et al.
O-acetyl sialic acid specific lectin from the crab Cancer antennarius
Methods Enzymol.
(1987) - et al.
A FTIR spectroscopy evidence of the interactions between wheat germ agglutinin and N-acetylglucosamine residues
Fed Eur Biochem Soc
(1999)
The lectins: carbohydrate binding proteins of plants and animals
Adv. Carbohydr. Chem. Biochem.
Purification and characterization of a natural agglutinin from the serum of the hermit crab Diogenes affinis
Biochim. Biophys. Acta
Naturally occurring bacterial agglutinin in the serum of the horseshoe crab Limulus polyphemus
J. Invertebr. Pathol.
Purification and characterization of a calcium independent lectin (PjLec) from the haemolymph of the shrimp Penaeus japornicus
Fish Shellfish Immunol.
Heterogeneous humoral and hemocyte associated lectins with N-acylaminosugar specificities from the blue crab, Callinectes sapidus Rathbun
Comp. Biochem. Physiol. B Comp. Biochem.
Purification and characterization of a lectin from Macrobrachium rosenbergii (Crustacea, Decapoda) hemolymph
Comp. Biochem. Physiol. Part B: Comp Biochem
Interaction of bovine erythrocytes N-glycolylneuraminic acid containing gangliosides and glycoproteins with a human Hanganutziu- deicher serum
Mol. Immunol.
Antibodies against 9-O-acetylated sialoglycans: a potent marker to monitor clinical status in childhood acute lymphoblastic leukemia
Clin. Biochem.
Introduction to lectins
Invertebrate lectins: distribution synthesis molecular biology and function
History of lectins: from hemaggultinins to biological recognition molecules
Glycobiology
Lectins, interconnecting proteins with biotechnological/pharmacological and therapeutic applications
Evidence – Based Complement Alt Med
Siglecs and their roles in the immune system
Nat. Rev. Immunol.
Neuraminidase- resistant sialic acid residues of gangliosides
Adv. Exp. Med. Biol.
An autosomal dominant gene regulates the extent of 9-O-acetylation of murine erythrocyte sialic acids. A probable explanation for the variation in capacity to activate the human alternate complement pathway
J. Exp. Med.
Purification and characterization of a sialic acid specific lectin from the hemolymph of the freshwater crab Paratelphusa jacquemontii
Eur. J. Biochem.
Sialic Acid Binding Lectin from the Hemolymph of an Estuarine Crab Metopograpsus messor (Forskal): Isolation, Purification, Characterization and Antibacterial Activity
Sialic acid specific lectins from Episesarma tetragonum (Decapoda, Grapsidae): isolation, purification and characterization
Int J Aquatic Biol
Purification and characterization of N-glycolyl neuraminic acid specific lectin from Scylla serrata
Eur. J. Biochem.
Cited by (7)
In vitro immune analysis of serum from the hemolymph of the anomuran crab Albunea symmysta (Linnaeus, 1758) displayed diversified reactions
2024, Journal of Invertebrate PathologySelective point-of-care detection of pathogenic bacteria using sialic acid functionalized gold nanoparticles
2021, TalantaCitation Excerpt :On the other hand, peaks at 1754 and 1722 cm−1 associated to ester carbonyl stretching modes are extremely weak and shifted to lower wavenumbers suggesting hydrogen bonding interactions as reported in the literature [25]. A broad band can be also observed in the 100-1500 cm−1 region due to SA C–O and C–OH stretching [26]. Thermogravimetric analysis of the SA-AuNPs (Supplementary material, Fig. S1a) revealed that after the degradation of the organic compound the remaining mass of gold represents 71.5 wt%, which implies a 28.5 wt% of SA.
Carbohydrate-Binding Activities of Agglutinins in Invertebrates from the Sea of Japan
2022, Russian Journal of Marine BiologyInvestigation of Lectins from Anomuran and Brachyuran Crabs
2022, Aquatic Lectins: Immune Defense, Biological Recognition and Molecular AdvancementsLectins in Penaeid Shrimps: Purification, Characterization, and Biological Significance
2022, Aquatic Lectins: Immune Defense, Biological Recognition and Molecular Advancements