Chemical Modification and Denaturation Effects on the Hemagglutinating Activity of Two Pterocarpus Species Seeds Lectins

Morenikeji Tolulope Folowosele

Biochemistry and Molecular Biology Department, Obafemi Awolowo University, Ile-Ife, Nigeria.

Oludele Olayemi Odekanyin *

Biochemistry and Molecular Biology Department, Obafemi Awolowo University, Ile-Ife, Nigeria.

Adenike Oluwaseun Adefila

Biochemistry Department, Faculty of Science, University of Ilesa, Ilesa, Nigeria.

Sinaola Praise Oyepitan

Biochemistry and Molecular Biology Department, Obafemi Awolowo University, Ile-Ife, Nigeria.

Eniola Racheal Owolabi

Biochemistry and Molecular Biology Department, Obafemi Awolowo University, Ile-Ife, Nigeria.

Ayomide Ifeoluwa Alobaloye

Biochemistry and Molecular Biology Department, Obafemi Awolowo University, Ile-Ife, Nigeria.

*Author to whom correspondence should be addressed.


Aims: Pterocarpus osun and Pterocarpus soyauxii seeds lectins were subjected to various chemical modifications in order to detect the amino acid residues involved in their hemagglutinating and sugar-binding activities.

Methodology: The lectins were purified using salt precipitation and size exclusion chromatography. Hemagglutinating activity and sugar specificity of the lectins were also established. Chemical modification of arginine was done using phenylglyoxal hydrate, and 5,5- dithiobis-(2-nitrobenzoic acid) (DTNB) was used to modify cysteine. Phenylmethylsulfonyl fluoride (PMSF) was employed for serine modification and tryptophan residue was modified with N-bromosuccinimide (NBS). Denaturants effects on the hemagglutinating activity were carried out with chaotropic agents, acid, disulphide bridge reducer and cross-linker agent.

Results: Pterocarpus osun seeds lectin is mannose specific while Pterocarpus soyauxii seeds lectin is galactose/lactose-binding lectin. Hemagglutinating activities of the two lectins were completely lost when tryptophan residue was modified with NBS and the loss was reversed by dialysis. Modifications of Cysteine, Arginine and Serine have no effect on the hemagglutinating activity of P. osun lectin. Nevertheless, the modifications of same amino acids slightly reduced the activity of P. soyauxii lectin, which dialysis and prolonged incubation were able to overturn. Mannose was found to bind and inhibit hemagglutinating activity of P. osun lectin in the presence of various modifiers but galactose and lactose could not inhibit the hemagglutinating activity of P. soyauxii lectin in the presence of modifiers. All denaturants employed significantly affected the hemagglutinating activity of the two lectins. However, the effects were reversible except for P. osun lectin denatured with 8M urea.

Conclusion: The results revealed that tryptophan residue is essential for hemagglutinating activity of the Pterocarpus species seeds lectins studied in this research. Cysteine, Arginine and Serine are also needed for sugar binding by P. soyauxii lectin but not so important in P. osun sugar binding ability.

Keywords: Lectin, amino acid, residue, denaturing agents, sugar, tryptophan, carbohydrate structure, arginine, glycoconjugate

How to Cite

Folowosele, M. T., Odekanyin , O. O., Adefila , A. O., Oyepitan , S. P., Owolabi, E. R., & Alobaloye , A. I. (2024). Chemical Modification and Denaturation Effects on the Hemagglutinating Activity of Two Pterocarpus Species Seeds Lectins. Chemical Science International Journal, 33(3), 89–99.


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Brooks S. Lectins as versatile tools to explore cellular glycosylation. Eur. J. Histochem. 2024;68(1):3959-3970

Katoch R, Tripathi A. Research advances and prospects of legume lectins. J. Biosci. 2021;46(4):104.

Roy R, Murphy PV, Gabius HJ. Multivalent carbohydrate-lectin interactions: How synthetic chemistry enables insights into nanometric recognition. Molecules. 2016; 21(5):629.

Santos AF, Da Silva MD, Napoleão TH, Paiva PM, Correia MD, Coelho LC. Lectins: Function, structure, biological properties and potential applications. Cur. Top. Pept. Prot. Res. 2014;15:41-62,

He M, Condict L, Richardson SJ, Brennan CS, Kasapis S. Molecular characterization of interactions between lectin-a protein from common edible mushroom (Agaricus bisporus)-and dietary carbohydrates. Food Hydrocolloids. 2024;146:109253.

Barre A, Bourne Y, Van Damme EJ, Rougé P. Overview of the structure–function relationships of mannose-specific lectins from plants, algae and fungi. Inter. J. Molec. Sci. 2019;20(2):254.

Boutureira O, Bernardes GJ. Advances in chemical protein modification. Chem. Rev. 2015;115(5):2174-95.

Schoellmann G, Shaw E. Direct evidence for the presence of histidine in the active center of chymotrypsin. Biochemistry. 1963;2(2):252-5.

Means GE, Feeney RE. Chemical modifications of proteins: history and applications. Bioconjug. Chem. 1990; 1(1):2-12

Chowdhury S, Ahmed H, Chatterjee BP. Chemical modification studies of Artocarpus lakoocha lectin artocarpin. Biochimie. 1991; 73(5):563-71.

DOI: 10.1016/0300-9084(91)90024-u PMID: 1764501.

Figueroa MOR, Lajolo FM. Effect of chemical modifications of Phaseolus vulgaris lectins on their biological properties. J. Agric Food Chem. 1997; 45(1): 639-43.

Hossain MA, Hossain MT, Absar N. Studies on the Chemical Modification of Mulberry Seed Lectin (MSL-1) and its Effect on Hemagglutinating Activity. Pak. J. Biol. Sci. 2006;9(4):674-680.

Loris R, Van Walle I, De Greve H, Beeckmans S, Deboeck F, Wyns L, Bouckaert J. Structural basis of oligomannose recognition by the Pterocarpus angolensis seed lectin. J. Molec. Biol. 2004;335(5):1227-40.

Dahat Y, Saha P, Mathew JT, Chaudhary SK, Srivastava AK, Kumar D. Traditional uses, phytochemistry and pharmacological attributes of Pterocarpus santalinus and future directions: A review. J. Ethnopharmacol. 2021;276:114127

Tchamadeu MC, Dzeufiet PD, Nana P, Nouga CK, Tsofack FN, Allard J, Blaes N, Siagat R, Zapfack L, Girolami JP, Tack I. Acute and sub-chronic oral toxicity studies of an aqueous stem bark extract of Pterocarpus soyauxii Taub (Papilionaceae) in rodents. J. Ethnopharmacol. 2011; 133(2):329-35.

Аrbаin D, Saputri GA, Syahputra GS, Widiyastuti Y, Susanti D, Taher M. Genus Pterocarpus: A review of ethnopharmacology, phytochemistry, biological activities, and clinical evidence. J. Ethnopharmacol. 2021;278:114316.

Chowdhury SR, Haldar S, Bhar R, Das S, Saha A, Pal K, Bandyopadhyay S, Paul J. Pterocarpus angolensis: Botanical, chemical and pharmacological review of an endangered medicinal plant of India. J. Exp. Biol. Agric. Sci. 2022;10:150-6.

Odekanyin OO, Akande OO. In-vitro Antioxidant and Antibacterial Potential of Mannose/Glucose-binding Pterocarpus osun Craib. Seeds Lectin. J. Appl. Life Sci. Internat. 2019;22(1):1-4

Odekanyin OO, Kayode AS, Adewuyi JO. Purification, characterization and antioxidant potential of a novel lectin from Pterocarpus soyauxii Taub seeds. Notulae Scientia Biologicae. 2019;11(1):112-21

Kuku A, Odekanyin OO, Okonji RE. Physicochemical properties of a lactose specific lectin from the seeds of Erythrina senegalensis DC. Ife J. Sci. 2012;14 (1):143-53.

Spande TT, Witkop B. Determination of the tryptophan content of proteins with N-bromosuccinimide. In Methods in Enzymology 1967; 11: 498-506. Academic Press.

Habeeb AF. Preparation of enzymically active water insoluble derivatives of trypsin. Arch. Biochem. Biophy. 1966; 119: 264-268.

Riordan JF. Arginyl residues and anion binding sites in proteins. Molec. Cellul. Biochem. 1979; 26(2):71-92.

Habeeb AF. Reaction of protein sulfhydryl groups with Ellman's reagent. In Methods in Enzymology Academic Press. 1972;25:457-464.

Pusam Y, Jaabir M, Jeyachandran S. Molecular Basis of Lectin–Carbohydrate Interaction. In: Elumalai, P., Lakshmi, S. (eds) Lectins. 2021. Springer, Singapore. Available:

Thakur A, Rana M, Lakhanpal TN, Ahmad A, Khan MI. Purification and characterization of lectin from fruiting body of Ganoderma lucidum: Lectin from Ganoderma lucidum. Biochim. Biophy Acta (BBA)-General Subjects. 2007;1770 (9):14 04-12.

Konozy EHE, Mulay R, Faca V, Ward RJ, Greene MJ, Roque-Barriera S, Sabharwal S. Bhide SV. Purification, some properties of a D-galactose-binding lectin from Erythrina indica and further characterization of seed lectin. Biochemistry. 2002;84:1035-1043.

Sikdar S, Chatterjee BP. Chemical modification studies on a blood group A-specific lectin, crotalarin (Crotalaria striata) and its effect on hemagglutinating activity. Molec. Cell. Biochem. 1990;96:107-16.

Datta PK, Figueroa MO, Lajolo FM. Chemical modification and sugar binding properties of two major lectins from Pinhao (Araucaria brasiliensis) seeds. J. Agric. Food Chem. 1993;41(11):1851-5.

Muramoto K, Kagawa D, Sato T, Ogawa T, Nishida Y, Kamiya H. Functional and structural characterization of multiple galectins from the skin mucus of conger eel, Conger myriaster. Comp Biochem Physiol B Biochem Mol Biol. 1999; 123(1):33-45.

Doi: 10.1016/s0305-0491(99)00037-1

Banerjee S, Naresh M, Swamy MJ. Effect of temperature and pH on the structure and stability of tumor-specific lectin jacalin and insights into the location of its tryptophan residues: CD, DSC and fluorescence studies. Inter. J. Biol. Macromol. 2024;260:129451.

Sultan NA, Kenoth R, Swamy MJ. Purification, physicochemical characterization, saccharide specificity, and chemical modification of a Gal/GalNAc specific lectin from the seeds of Trichosanthes dioica. Arch. Biochem. Biophy. 2004;432(2):212-21.

Rasmussen A, Rasmussen T, Edwards MD, Schauer D, Schumann U, Miller S, Booth IR. The role of tryptophan residues in the function and stability of the mechanosensitive channel MscS from Escherichia coli. Biochemistry. 2007; 46(38):10899-908. DOI: 10.1021/bi701056k

Betts MJ, Russell RB. Amino acid properties and consequences of substitutions. Bioinformatics for Geneticists. 2003;317: 289.

Yeasmin T, Kashem T, Pazzaque A, Absar N. Purification and characterization of 3 galactose-specific lectins from mulberry seeds (Morus spp.). Eur. J. Biochem. 2001;268:6005-6010.

Singh P, Saxona K. Effect of temperature, pH and denaturing agents on biological activity of MCJ lectins. Chemi. Sci. Transit. 2013;2(4):1508-1512.

Poole LB, Furdui CM, King SB. Introduction to approaches and tools for the evaluation of protein cysteine oxidation. Essays Biochem. 2020;64(1):1-17.

DOI: 10.1042/EBC20190050

Canchi DR, Paschek D, García AE. Equilibrium study of protein denaturation by urea. J. Americ. Chem. Soc. 2010; 132(7):2338-44.

Wingfield PT, editor. Use of protein folding reagents. Current Protocols in Protein Science. 2016; 84(1):A-3A.

Mullaiselvan I, Nelluri P, Nisha B, Saha D. Properties and Applications of Proteins from Pseudocereals. Pseudocereals: Production, Processing, and Nutrition. 2024;132.

Cao L, Rantwijk F, Sheldon R. Cross-linked enzyme aggregates: a simple and effective method for the immobilization of penicillin acylase. Organic Letters. 2000; 2(10):1361-1364.