Characterization of O-glycan binding lectin from the red alga Hydropuntia eucheumatoides
Keywords:Carbohydrate binding specificity, Hydropuntia eucheumatoides, lectin, molecular mass, red alga, stable
The red alga, Hydropuntia eucheumatoides is one of the algal genera from which agar is commercially extracted, and is the main source of agar in the world. The lectin HEL from the red alga H. eucheumatoides was isolated by a combination of aqueous ethanol extraction, ethanol precipitation, ion exchange and filtration chromatography. Lectin gave a single band with molecular mass of 17,000 Da in both non-reducing and reducing SDS-PAGE conditions, therefore lectin exists in monomeric form. The hemagglutination activities of HEL were stable over a wide range of pH from 3 to 10, temperature up 60 oC and not affected by either the presence of EDTA or addition of divalent cations, indicating that lectin requires no metal for biological activity. The hemagglutination activities of HEL were not inhibited by monosaccharides and glycoproteins, D-glucose, D-mannose, D-galactose, D-xylose, N-acety-D-mannosamine, transferin, fetuin and yeast mannan, but strongly inhibited by monosaccharides containing acetamido groups at equatorial C2 position, such as N-acetyl-galactosamine, N-acetyl-glucosamine, N-acetyl-neuraminic acid and glycoprotein porcine stomach mucin bearing O-glycans. Thus, lectin is specific for O-glycans and may recognize the sequences GalNAcαSer/Thr, GalNAc(α1-3)[Fuc(α1-2)]Gal(β1-4)GlcNAc(β1-3)GalNAc- and GluNAc(α1-4)Gal- under interacting with the acetamido groups at equatorial C2 position of the terminal sugar residues in oligosaccharide structures of O-glycans. The red alga H. eucheumatoides could promise to be a source of valuable lectins for application in biochemistry and biomedicine.
Ainouz IL, Sampaio AH, Benevides NMB, Freitas ALP, Costa FHF, Carvalho MC, Pinheiro-Joventino F (1992) Agglutination of enzyme treated erythrocytes by Brazilian marine algae. Bot Mar 35: 475–479.
Chiles TC, Bird KT (1989) A comparative study of the animal erythrocyte agglutinins from marine algae. Comp Biochem Physiol 94B: 107–111.
Chiles TC, Bird KT (1990) Gracilaria tikvahiae agglutinin. Partial purification and preliminary characterization of its carbohydrate specificity. Carbohydr Res 207: 319-326.
Dinh Thanh Trung, Vo Thi Dieu Trang, Ngo Thi Duy Ngoc, Phan Thi Hoai Trinh, Tran Thi Hai Yen, Le Dinh Hung (2017) Haemagglutination activity of the extracts from some Vietnam marine invertebrates. Vietnam J Biotechnol 15: 691-701.
Freitas ALP, Teixeira DIA, Costa FHF, Farias WRL, Lobato ASC, Sampaio AH, Benevides NMB (1997) A new survey of Brazilian marine algae for agglutinins. J Appl Phycol 9: 495–501.
Hirayama M, Shibata H, Imamura K, Sakaguchi T, Hori K (2016) High-mannose specific lectin and its recombinants from a carrageenophyta Kappaphycus alvarezii represent a potent anti-HIV activity through high-affinity binding to the viral envelope glycoprotein gp120. Mar Biotechnol 18:144-160.
Hori K, Oiwa C, Miyazawa K, Ito K (1988) Evidence for wide distribution of agglutinins in marine algae. Bot Mar 31: 133–138.
Hori K, Miyazawa K, Ito K (1990) Some common properties of lectins from marine algae. Hydrobiologia 204/205: 561-566.
Hori K, Sato Y, Ito K, Fujiwara Y, Iwamoto Y, Makino H, Kawakubo A (2007) Strict specificity for high mannose-type N-glycans and primary structure of a red alga Eucheuma serra lectin. Glycobiology 17: 479-491.
Huynh Quang Nang, Nguyen Huu Dinh (1998) The seaweed resources of Vietnam. In: Critchley AT, Ohno M (eds) Seaweed resources of the World. JICA, Japan pp: 62-69.
Kakita H, Fukuoka S, Obika H, Kamishima H (1999) Isolation and characterisation of a fourth hemagglutinin from the red alga, Gracilaria verrucosa, from Japan. J Appl Phycol 11: 49-56.
Karlsson NG, Nordman H, Karlsson H, Carlstedt I, Hansson GC (1997) Glycosylation differences between pig gastric mucin populations: A comparative study of the neutral oligosaccharides using mass spectrometry. J Biochem 326: 911-917.
Kawakubo A, Makino H, Ohnishi J, Hirohara H, Hori K (1997) The marine red alga Eucheuma serra J. Agardh, a high yielding source of two isolectins. J Appl Phycol 9: 331-338.
Kawakubo A, Makino H, Ohnishi J, Hirohara H, Hori K (1999) Occurrence of highly yielded lectins homologous within the genus Eucheuma. J Appl Phycol 11: 149-156.
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.
Le Dinh Hung, Hori K, Huynh Quang Nang (2009a) Screening and preliminary characterization of hemagglutinins in Vietnamese marine algae. J Appl Phycol 21: 89-97.
Le Dinh Hung, Sato T, Shibata H, Hori K (2009b) Biochemical comparison of lectins among three different color strains of the red alga Kappaphycus alvarezii. Fish Sci 75: 723-730.
Le Dinh Hung, Hori K, Huynh Quang Nang, Tran Kha, Le Thi Hoa (2009c) Seasonal changes in growth rate, carrageenan yield and lectin content in the red alga Kappaphycus alvarezii cultivated in Camranh Bay, Vietnam. J Appl Phycol 21:265-272.
Le Dinh Hung, Sato Y, Hori K (2011) High-mannose N-glycan-specific lectins from the red alga Kappaphycus striatum (Carrageenophyte). Phytochemistry 72: 855-861.
Le Dinh Hung, Bui Minh Ly, Vo Thi Dieu Trang, Ngo Thi Duy Ngoc, Le Thi Hoa, Phan Thi Hoai Trinh (2012) A new screening for hemagglutinins from Vietnamese marine macroalgae. J Appl Phycol 24: 227- 235.
Le Dinh Hung, Ngo Thi Duy Ngoc, Hori K (2013) Isolation and characterization of novel lectin from the red alga Gracilaria salicornia. Vietnam J Biotechnol 11: 743-753.
Le Dinh Hung, Hirayama M, Bui Minh Ly, Hori K (2015a) Purification, primary structure, and biological activity of high-mannose N-glycan-specific lectin from the cultivated Eucheuma denticulatum. J Appl Phycol 27: 1657-1669.
Le Dinh Hung, Hirayama M, Bui Minh Ly, Hori K (2015b) Biological activity, cDNA cloning and primary structure of lectin KSA-2 from the cultivated red alga Kappaphycus striatum (Schmitz) Doty ex Silva. Phytochem Lett 14: 99-105.
Le Dinh Hung, Hirayama M, Hori K (2016) Cloning and characterizing cDNA sequences coding high-mannose N-glycan binding lectins from cultivated red algae Eucheuma denticulatum and Kappaphycus striatum. Vietnam J Biotechnol 14: 327-336.
Le Dinh Hung, Bui Minh Ly, Vo Thi Hao, Dinh Thanh Trung, Vo Thi Dieu Trang, Phan Thi Hoai Trinh, Ngo Thi Duy Ngoc, Thai Minh Quang (2018a) Purification, characterization and biological effect of lectin from the marine sponge Stylissa flexibilis (Lévi, 1961). Comp Biochem Physiol, Part B 216: 32-38.
Le Dinh Hung, Le Thi Hoa, Le Nhu Hau, Dinh Thanh Trung (2018b) The lectin accumulation, growth rate, carrageenan yield, and quality from the red alga Kappaphycus striatus cultivated at Camranh Bay, Vietnam. J Appl Phycol
Leite YFMM, Silva LMCM, Amorim RCN, Freire EA, Jorge DMM, Grangeiro TB, Benevides NMB (2005) Purification of a lectin from the marine red alga Gracilaria ornata and its effect on the development of the cowpea weevil Callosobruchus maculatus (Coleoptera: Bruchidae). Biochim Biophys Acta 1724: 137-145.
Lima MEP, Carneiro ME, Nascimento AE, Grangeiro TB, Holanda ML, Amorim RCN, Benevides NMB (2005) Purification of a lectin from the marine red alga Gracilaria cornea and its effects on the Cattle Tick Boophilus microplus (Acari: Ixodidae). J Agric Food Chem 53: 6414-6419.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951)
Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265-275.
Mori T, O’Keefe BR, Sowder RC, Bringans S, Gardella RS, Berg S, Cochran P, Turpin JA, Buckheit RW, McMahon JBJr, Boyd MR (2005) Isolation and characterization of griffithsin, a novel HIV-inactivating protein, from the red alga Griffithsia sp. J Biol Chem 280: 9345–9353.
Mu J, Hirayama M, Sato Y, Morimoto K, Hori K (2017) A novel high-mannose specific lectin from the green alga Halimeda renschii exhibits a potent anti-influenza virus activity through high-affinity binding to the viral hemagglutinin. Mar Drugs 15, 255. Doi: 10.3390/md15080255.
Nagano CS, Debray H, Nascimento KS, Pinto VPT, Cavada BS, Saker-Sampaio S, Farias WRL, Sampaio AH, Calvete JJ (2005) HCA and HML isolated from the red marine algae Hypnea cervicornis and Hypnea musciformis define a novel lectin family. Protein Sci 14: 2167-2176.
Okamoto R, Hori K, Miyazawa K, Ito K (1990) Isolation and characterization of a new hemagglutinin from the red alga Gracilaria bursa-pastoris. Experientia 46: 975-977.
Oliveira EC, Alveal K, Anderson JR (2000) Mariculture of the agar-producing gracilarioid red algae. Rev Fish Sci 8: 345-377.
Rogers DJ, Hori K (1993) Marine algal lectins: new developments. Hydrobiologia 260/261: 589-593.
Sato Y, Morimoto K, Hirayama M, Hori K (2011a) High mannose-specific lectin (KAA-2) from the red alga Kappaphycus alvarezii potently inhibits influenza virus infection in a strain-independent manner. Biochem Biophys Res Com 405: 291-296.
Sato Y, Hirayama M, Morimoto K, Yamamoto N, Okuyama S, Hori K (2011b) High mannose-binding lectin with preference for the cluster of α1–2-mannose from the green alga Boodlea coacta is a potent entry inhibitor of HIV-1 and influenza viruses. J Biol Chem 286: 19446-19458.
Sato Y, Morimoto K, Kubo T, Sakaguchi T, Nishizono A, Hirayama M, Hori K (2015) Entry inhibition of influenza viruses with high mannose binding lectin ESA-2 from the red alga Eucheuma serra through the recognition of viral hemagglutinin. Mar Drugs 13: 3454-3465.
Sharon N, Lis H (2003) Lectins (second edition), Kluwer Academic Publishers, The Netherlands.
Tsuji T, Yamamoto K, Irimura T, Osawa T (1981) Structure of carbohydrate unit A of porcine thyroglobulin. J Biochem 195: 691-699.
Van Halbeek H, Dorland L, Vliegenthart JFG, Kochetkov K, Arbatsky NP, Derevitskaya VA (1982) Characterization of the primary structure and the microheterogeneity of the carbohydrate chains of porcine blood-group H substance by 500-MHz 1H-NMR spectroscopy. Eur J Biochem 127: 21-29.
Wopereis S, Lefeber DJ, Morava E, Wevers RA (2006) Mechanisms in protein O-glycan biosynthesis and clinical and molecular aspects of protein O-glycan biosynthesis defects: A Review. Clinical Chemistry 52: 574-600.
Xiong C, Li W, Liu H, Zhang W, Dou J, Bai X, Du Y, Ma X (2006) A normal mucin-binding lectin from the sponge Craniella australiensis. Comp Biochem Physiol, Part C 143: 9–16.
Yamamoto K, Tsuji T, Irimura T, Osawa T (1981) The structure of carbohydrate unit B of porcine thyroglobulin. J Biochem 195: 701-713.