Purification and Partial Characterization of Agglutinin Lectin from Heamolymph of German Cockroach, Blattella germanica
Abstract
Background: Lectin molecules have crucial biological role in insects’ immune system. The aim of present study was to find the agglutinin activities in haemolymph of German cockroach, Belatella germanica with appropriate screening and purification.
Methods: The heamolymph of cockroach was collected and agglutinin test performed against different animal and human red blood cells (RBC). Then sugar inhibition assay was carried out to find carbohydrate specific binding lec¬tin. The proteins of haemolymph was purified using ion-exchange chromatography (HPLC) and each fraction was tested for agglutinin activity. Finally the molecular weight of the agglutinin protein was determined using SDS-page.
Results: The most agglutinin activity of haemolymph was found against RBC of mouse at titer 1/128ml/L dilution and sugar inhibition assay showed that fucos, N-acetyglucoseamine and galactose reduced titer of agglutinin to ½ml/L. Only one fraction of heamolymph at rotation time of 36 minute showed agglutinin activity. The molecular weight of this lectin was measured as 120Kds.
Conclusion: The range of agglutinin activities against different RBC indicates that the isolated lectin is not specific for a particular carbohydrate. In addition, the isolated lectin at low concentration present in heamolymph should be an innate lactin not secreted, because we found it without any trigger immunity of the insect.
1. Arumugam G, Sreeramulu B, Paulchamy R, Thangavel S, Sundaram J (2017) Purification and functional character¬ization of lectin with phenoloxidase activity from the hemolymph of cock¬roach, Periplaneta americana. Arch Insect Biochem Physiol. 95(2): e21390.
2. Basseri HR, Doosti S, Akbarzadeh K, Nateghpour M, Whitten MM, Ladoni H (2008) Competency of Anopheles stephensi mysorensis strain for Plas¬modium vivax and the role of in¬hibi¬tory carbohydrates to block its sporo¬gonic cycle. Mal J. 7: 131.
3. Basseri HR, Emmami, N, Haji-Hosseini R, Abolhasani M, Moradi A (2008) Biological transmission of bacteria in¬hibit by hemolymph lectins of Amer¬ican cockroach. Iranian J Pub¬lic Health. 37: 75–82.
4. Basseri HR, Mohamadzadeh Hajipirloo H, Mohammadi Bavani M, Whitten MM (2013) Comparative susceptibil¬ity of different biological forms of Anophe¬les stephensi to Plasmodium berghei anka strain. Plos One. 8: E75413.
5. Bi J, Feng F, Li J, Mao J, Ning M, Song X, Xie J, Tang J, Li B (2019) A c-type lectin with a single carbohy-drate-recognition domain involved in the innate immune response of Tri¬bolium castaneum. Insect Mol Biol. 28(5): 649–661.
6. Bradford MM (1976) A rapid and sensi-tive method for the quantitation of mi¬crogram quantities of protein uti-lizing the principle of protein-dye bind-ing. Anal Biochem. 72: 248–254.
7. Bulgakov AA, Park KI, Choi KS, Lim HK, Cho M (2004) Purification and char¬acterization of a lectin isolated from the manila clam Ruditapes phil-ippinar¬um in Korea. Fish Shellfish Immu¬nol. 16: 487–499.
8. Chen C, Durrant HJ, Newton RP, Ratcliffe NA (1995) A study of nov-el lectins and their involvement in the activa¬tion of the prophenoloxi-dase system in Blaberus discoidalis. Biochem J. 310(1): 23–31.
9. Chen C, Ratcliffe NA, Rowley AF (1993) Detection, isolation and characteriza¬tion of multiple lectins from the haemo¬lymph of the cockroach Blaberus dis¬coidalis. Biochem J. 294(1): 181–190.
10. Chen C, Rowley AF, Newton RP, Ratcliffe NA (1999) Identification, purifica-tion and properties of a beta-1,3-glucan-specific lectin from the serum of the cockroach, Blaberus discoidalis which is implicated in immune defence re¬actions. Comp Biochem Physiol B Bi¬ochem Mol Biol. 122: 309–319.
11. Do DC, Zhao Y, Gao P (2016) Cock-roach allergen exposure and risk of asthma. Allergy. 71: 463–474.
12. Drickamer K, Taylor ME (1993) Biolo-gy of animal lectins. Annu Rev Cell Biol. 9: 237–264.
13. Gül N, Ayvali C (2002) Purification and determination of the molecular struc¬ture of hemolymph lectin of Agrotis segetum (Denis and Schiff). Turk J Biol. 26: 49–55.
14. Kawasaki K, Kubo T, Natori S (1993) A novel role of Periplaneta lectin as an opsonin to recognize 2-keto-3-deoxy octonate residues of bacterial lipo¬pol¬ysaccharides. Comp Biochem Phys¬iol B. 106: 675–680.
15. Kawasaki K, Kubo T, Natori S (1996) Presence of the Periplaneta lectin-re¬lated protein family in the ameri-can cockroach Periplaneta america-na. In¬sect Biochem Mol Biol. 26: 355–364.
16. Kubo T, Kawasaki K, Natori S (1993) Transient appearance and localiza-tion of a 26-kda lectin, a novel mem¬ber of the Periplaneta lectin family, in regenerating cockroach leg. Dev Biol. 156: 381–390.
17. Kubo T, Natori S (1987) Purification and some properties of a lectin from the hemolymph of Periplaneta amer¬icana (american cockroach). Eur J Bio¬chem. 168: 75–82.
18. Lackie AM, Vasta GR (1988) The role of galactosyl-binding lectin in the cel¬lular immune response of the cock¬roach Periplaneta americana (Dic¬tyoptera). Immunology. 64: 353–357.
19. Lam SK, Ng TB (2011) Lectins: produc-tion and practical applications. Appl Microbiol Biot. 89: 45–55.
20. Makela O (1957) Studies in hemaggluti-nins of leguminosae seeds. Ann Med Exp Biol Fenn. 35: 1–133.
21. Olafsen JA (1995) Lectins: models of natural and induced molecules in in-vertebrates. In: Cooper EL (Ed) Ad-vances in Comparative Environmen-tal Physiology. Springer-Verlag, Ber¬lin, pp. 49–76.
22. Ottaviani E, Malagoli D, Franchini A (2004) Invertebrate humoral factors: cytokines as mediators of cell sur-vival. Prog Mol Subcell Biol. 34: 1–25.
23. Ratcliffe A, Fryer PR, Hardingham TE (1985) Proteoglycan biosynthesis in chondrocytes: protein a-gold locali-zation of proteoglycan protein core and chondroitin sulfate within golgi sub compartments. J Cell Biol. 101: 2355–2365.
24. Schnitger AK, Yassine H, Kafatos FC, Osta MA (2009) Two c-type lectins cooperate to defend Anopheles gam-biae against gram-negative bacteria. J Biol Chem. 284: 17616–17624.
25. Thakur K, Kaur T, Kaur M, Hora R, Singh J (2019) Exploration of carbo-hydrate binding behavior and anti-pro¬lif¬erative activities of Arisaema tortu¬osum lectin. BMC Mol Biol. 20: 15.
26. Viswambari Devi R, Basilrose MR, Mer¬cy PD (2010) Prospect for lectis in arthropods. Ital J Zool. 3: 257–260.
27. Ward SE, O'Sullivan JM, Drakeford C, Aguila S, Jondle CN, Sharma J, Fal-lon PG, Brophy TM, Preston RJS, Smyth P, Sheils O, Chion A, O'Don-nell JS (2018) A novel role for the macrophage galactose-type lectin re¬ceptor in mediating von willebrand factor clearance. Blood. 131: 911–916.
28. Wilson R, Chen C, Ratcliffe NA (1999) Innate immunity in insects: the role of multiple, endogenous serum lec-tins in the recognition of foreign in-vaders in the cockroach, Blaberus discoidalis. J Immunol. 162: 1590–1596.
29. Xia X, You M, Rao XJ, Yu XQ (2018) Insect c-type lectins in innate im-mun¬ity. Dev Comp Immunol. 83: 70–79.
30. Yu XQ, Kanost MR (2000) Immulectin-2, a lipopolysaccharide-specific lec-tin from an insect, Manduca sexta, is induced in response to gram-negative bacteria. J Biol Chem. 275: 37373–37381.
31. Yu XQ, Kanost MR (2003) Manduca sexta lipopolysaccharide-specific im-mulec¬tin-2 protects larvae from bacterial in¬fection. Dev Comp Im-munol. 27: 189–196.
32. Zheng X, Xia Y (2012) Beta-1, 3-glucan recognition protein (betagrp) is es-sen¬tial for resistance against fungal path¬ogen and opportunistic patho-genic gut bacteria in Locusta migra-toria ma¬nilensis. Dev Comp Immu¬nol. 36: 602–609.
2. Basseri HR, Doosti S, Akbarzadeh K, Nateghpour M, Whitten MM, Ladoni H (2008) Competency of Anopheles stephensi mysorensis strain for Plas¬modium vivax and the role of in¬hibi¬tory carbohydrates to block its sporo¬gonic cycle. Mal J. 7: 131.
3. Basseri HR, Emmami, N, Haji-Hosseini R, Abolhasani M, Moradi A (2008) Biological transmission of bacteria in¬hibit by hemolymph lectins of Amer¬ican cockroach. Iranian J Pub¬lic Health. 37: 75–82.
4. Basseri HR, Mohamadzadeh Hajipirloo H, Mohammadi Bavani M, Whitten MM (2013) Comparative susceptibil¬ity of different biological forms of Anophe¬les stephensi to Plasmodium berghei anka strain. Plos One. 8: E75413.
5. Bi J, Feng F, Li J, Mao J, Ning M, Song X, Xie J, Tang J, Li B (2019) A c-type lectin with a single carbohy-drate-recognition domain involved in the innate immune response of Tri¬bolium castaneum. Insect Mol Biol. 28(5): 649–661.
6. Bradford MM (1976) A rapid and sensi-tive method for the quantitation of mi¬crogram quantities of protein uti-lizing the principle of protein-dye bind-ing. Anal Biochem. 72: 248–254.
7. Bulgakov AA, Park KI, Choi KS, Lim HK, Cho M (2004) Purification and char¬acterization of a lectin isolated from the manila clam Ruditapes phil-ippinar¬um in Korea. Fish Shellfish Immu¬nol. 16: 487–499.
8. Chen C, Durrant HJ, Newton RP, Ratcliffe NA (1995) A study of nov-el lectins and their involvement in the activa¬tion of the prophenoloxi-dase system in Blaberus discoidalis. Biochem J. 310(1): 23–31.
9. Chen C, Ratcliffe NA, Rowley AF (1993) Detection, isolation and characteriza¬tion of multiple lectins from the haemo¬lymph of the cockroach Blaberus dis¬coidalis. Biochem J. 294(1): 181–190.
10. Chen C, Rowley AF, Newton RP, Ratcliffe NA (1999) Identification, purifica-tion and properties of a beta-1,3-glucan-specific lectin from the serum of the cockroach, Blaberus discoidalis which is implicated in immune defence re¬actions. Comp Biochem Physiol B Bi¬ochem Mol Biol. 122: 309–319.
11. Do DC, Zhao Y, Gao P (2016) Cock-roach allergen exposure and risk of asthma. Allergy. 71: 463–474.
12. Drickamer K, Taylor ME (1993) Biolo-gy of animal lectins. Annu Rev Cell Biol. 9: 237–264.
13. Gül N, Ayvali C (2002) Purification and determination of the molecular struc¬ture of hemolymph lectin of Agrotis segetum (Denis and Schiff). Turk J Biol. 26: 49–55.
14. Kawasaki K, Kubo T, Natori S (1993) A novel role of Periplaneta lectin as an opsonin to recognize 2-keto-3-deoxy octonate residues of bacterial lipo¬pol¬ysaccharides. Comp Biochem Phys¬iol B. 106: 675–680.
15. Kawasaki K, Kubo T, Natori S (1996) Presence of the Periplaneta lectin-re¬lated protein family in the ameri-can cockroach Periplaneta america-na. In¬sect Biochem Mol Biol. 26: 355–364.
16. Kubo T, Kawasaki K, Natori S (1993) Transient appearance and localiza-tion of a 26-kda lectin, a novel mem¬ber of the Periplaneta lectin family, in regenerating cockroach leg. Dev Biol. 156: 381–390.
17. Kubo T, Natori S (1987) Purification and some properties of a lectin from the hemolymph of Periplaneta amer¬icana (american cockroach). Eur J Bio¬chem. 168: 75–82.
18. Lackie AM, Vasta GR (1988) The role of galactosyl-binding lectin in the cel¬lular immune response of the cock¬roach Periplaneta americana (Dic¬tyoptera). Immunology. 64: 353–357.
19. Lam SK, Ng TB (2011) Lectins: produc-tion and practical applications. Appl Microbiol Biot. 89: 45–55.
20. Makela O (1957) Studies in hemaggluti-nins of leguminosae seeds. Ann Med Exp Biol Fenn. 35: 1–133.
21. Olafsen JA (1995) Lectins: models of natural and induced molecules in in-vertebrates. In: Cooper EL (Ed) Ad-vances in Comparative Environmen-tal Physiology. Springer-Verlag, Ber¬lin, pp. 49–76.
22. Ottaviani E, Malagoli D, Franchini A (2004) Invertebrate humoral factors: cytokines as mediators of cell sur-vival. Prog Mol Subcell Biol. 34: 1–25.
23. Ratcliffe A, Fryer PR, Hardingham TE (1985) Proteoglycan biosynthesis in chondrocytes: protein a-gold locali-zation of proteoglycan protein core and chondroitin sulfate within golgi sub compartments. J Cell Biol. 101: 2355–2365.
24. Schnitger AK, Yassine H, Kafatos FC, Osta MA (2009) Two c-type lectins cooperate to defend Anopheles gam-biae against gram-negative bacteria. J Biol Chem. 284: 17616–17624.
25. Thakur K, Kaur T, Kaur M, Hora R, Singh J (2019) Exploration of carbo-hydrate binding behavior and anti-pro¬lif¬erative activities of Arisaema tortu¬osum lectin. BMC Mol Biol. 20: 15.
26. Viswambari Devi R, Basilrose MR, Mer¬cy PD (2010) Prospect for lectis in arthropods. Ital J Zool. 3: 257–260.
27. Ward SE, O'Sullivan JM, Drakeford C, Aguila S, Jondle CN, Sharma J, Fal-lon PG, Brophy TM, Preston RJS, Smyth P, Sheils O, Chion A, O'Don-nell JS (2018) A novel role for the macrophage galactose-type lectin re¬ceptor in mediating von willebrand factor clearance. Blood. 131: 911–916.
28. Wilson R, Chen C, Ratcliffe NA (1999) Innate immunity in insects: the role of multiple, endogenous serum lec-tins in the recognition of foreign in-vaders in the cockroach, Blaberus discoidalis. J Immunol. 162: 1590–1596.
29. Xia X, You M, Rao XJ, Yu XQ (2018) Insect c-type lectins in innate im-mun¬ity. Dev Comp Immunol. 83: 70–79.
30. Yu XQ, Kanost MR (2000) Immulectin-2, a lipopolysaccharide-specific lec-tin from an insect, Manduca sexta, is induced in response to gram-negative bacteria. J Biol Chem. 275: 37373–37381.
31. Yu XQ, Kanost MR (2003) Manduca sexta lipopolysaccharide-specific im-mulec¬tin-2 protects larvae from bacterial in¬fection. Dev Comp Im-munol. 27: 189–196.
32. Zheng X, Xia Y (2012) Beta-1, 3-glucan recognition protein (betagrp) is es-sen¬tial for resistance against fungal path¬ogen and opportunistic patho-genic gut bacteria in Locusta migra-toria ma¬nilensis. Dev Comp Immu¬nol. 36: 602–609.
| Files | ||
| Issue | Vol 14 No 2 (2020) | |
| Section | Original Article | |
| DOI | https://doi.org/10.18502/jad.v14i2.3732 | |
| Keywords | ||
| German cockroach; Heamolymph; Lectin | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Nabavi Z, Baniardalani M, Basseri H. Purification and Partial Characterization of Agglutinin Lectin from Heamolymph of German Cockroach, Blattella germanica. J Arthropod Borne Dis. 2020;14(2):144–152.
