Purification and characterization of a heteromultimeric glycoprotein from Artocarpus heterophyllus latex with an inhibitory effect on human blood coagulation
Plant latex has many health benefits and has been used in folk medicine. In this study, the biological effect of Artocarpus heterophyllus (jackfruit) latex on human blood coagulation was investigated. By a combination of heat precipitation and ion-exchange chromatography, a heat stable heteromultimeric glycoprotein (HSGPL1) was purified from jackfruit milky latex. The apparent molecular masses of the monomeric proteins on SDS/PAGE were 33, 31 and 29 kDa. The isoelectric points (pIs) of the monomers were 6.63, 6.63 and 6.93, respectively. Glycosylation and deglycosylation tests confirmed that each subunit of HSGPL1 formed the native multimer by sugar-based interaction. Moreover, the multimer of HSGPL1 also resisted 2-mercaptoethanol action. Peptide mass fingerprint analysis indicated that HSGPL1 was a complex protein related to Hsps/chaperones. HSGPL1 has an effect on intrinsic pathways of the human blood coagulation system by significantly prolonging the activated partial thrombin time (APTT). In contrast, it has no effect on the human extrinsic blood coagulation system using the prothrombin time (PT) test. The prolonged APTT resulted from the serine protease inhibitor property of HSGPL1, since it reduced activity of human blood coagulation factors XIa and α-XIIa.
- Ajayi IA (2008) Comparative study of the chemical composition and mineral element content of Artocarpus heterophyllus and Treculia africana seeds and seed oils. Bioresour Technol 99: 5125-5129.
- Bode W, Huber R (2000) Structural basis of the endoproteinase-protein inhibitor interaction. Biochim Biophys Acta 1477: 241-252.
- Bohlooli S, Mohebipoor A, Mohammadi S, Kouhnavard M, Pashapoor S (2007) Comparative study of fig tree efficacy in the treatment of common warts (Verruca vulgaris) vs. cryotherapy. Int J Dermatol 46: 524-526.
- Brown BA (1988) Haematology: Principles and Procedures. pp 195-215. Lea and Febiger, Philadelphia.
- Carranco R, Almoguera C, Jordano J (1997) A plant small heat shock protein gene expressed during zygotic embryogenesis but noninducible by heat stress. J Biol Chem 272: 27470-27475.
- Chen LL, Rosa JJ, Turner S, Pepinsky RB. (1991) Production of multimeric forms of CD4 through a sugar-based cross-linking strategy. J Biol Chem 266: 18237-18243.
- Dafforn TR, Della M, Miller AD (2001) The molecular interactions of heat shock protein 47 (Hsp47) and their implications for collagen biosynthesis. J Biol Chem 276: 49310-49319.
- Fernando MR, Wickramasinghe N, Thabrew MI, Ariyananda PL, Karunanayake EH (1991) Effect of Artocarpus heterophyllus and Asteracanthus longifolia on glucose tolerance in normal human subjects and in maturity-onset diabetic patients. J Ethnopharmacol 31: 277-282.
- Fonseca KC, Morais NC, Queiroz MR, Silva MC, Gomes MS, Costa JO, Mamede CC, Torres FS, Penha-Silva N, Beletti ME, Canabrava HA, Oliveira F (2010) Purification and biochemical characterization of Eumiliin from Euphorbia milii var. hislopii latex. Phytochemistry 71: 708-715.
- Gurung S, Skalko-Basnet N (2009) Wound healing properties of Carica papaya latex: in vivo evaluation in mice burn model. J Ethnopharmacol 121: 338-341.
- Habeeb AF (1996) Determination of free amino groups in proteins by trinitrobenzenesulfonic acid. Anal Biochem 14: 328-336.
- Hatakeyama T, Kohzaki H, Yamasaki N (1992) A microassay for proteases using succinylcasein as a substrate. Anal Biochem 204: 181-184.
- Hayashi K, Takehisa T, Hamato N, Takano R, Hara S, Miyata T, Kato H (1994) Inhibition of serine proteases of the blood coagulation system by squash family protease inhibitors. J Biochem 116: 1013-1018.
- Hernández MA, Avila J, Andreu JM (1986) Physicochemical characterization of the heat-stable microtubule-associated protein MAP2. Eur J Biochem 154: 41-48.
- Ivanciu L, Gerard RD, Tang H, Lupu F, Lupu C (2007) Adenovirus-mediated expression of tissue factor pathway inhibitor-2 inhibits endothelial cell migration and angiogenesis. Arterioscler Thromb Vasc Biol 27: 310-316.
- Jethmalani SM, Henle KJ (1998) Interaction of heat stress glycoprotein GP50 with classical heat-shock proteins. Exp Cell Res 239: 23-30.
- Kabir S (1995) The isolation and characterisation of jacalin [Artocarpus heterophyllus (jackfruit) lectin] based on its charge properties. Int J Biochem Cell Biol 27: 147-156.
- Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.
- Maier W, Meier B (2002) Mechanical Treatment to Stabilize Plaque. In Mechanical Treatment to Stabilize Plaque. Brown DL, pp 401-426. Marcel Dekker, New York, London.
- Mekkriengkrai D, Ute K, Swiezewska E, Chojnacki T, Tanaka Y, Sakdapipanich JT (2004) Structural characterization of rubber from jackfruit and euphorbia as a model of natural rubber. Biomacromolecules 5: 2013-1019.
- Osoniyi O, Onajobi F (2003) Coagulant and anticoagulant activities in Jatropha curcas latex. J Ethnopharmacol 89: 101-105.
- Pasamontes L, Haiker M, Wyss M, Tessier M, van Loon AP (1997) Gene cloning, purification, and characterization of a heat-stable phytase from the fungus Aspergillus fumigatus. Appl Environ Microbiol 63: 1696-1700.
- Prasad KMR, Virupaksha TK (1990) Purification and characterization of a protease from jackfruit latex. Phytochemistry 29: 1763-1766.
- Shevchenko A, Wilm M, Vorm O, Mann M (1996) Mass spectrometric sequencing of proteins from silver-stained polyacrylamide gels. Anal Chem 68: 850-858.
- Shih MD, Hsieh TY, Lin TP, Hsing YI, Hoekstra FA (2010) Characterization of two soy bean (Glycine max L.) LEA IV proteins by circular dichroism and Fourier transform infrared spectrometry. Plant Cell Physiol 51: 395-407.
- Shimokawa K, Katayama M, Matsuda Y, Takahashi H, Hara I, Sato H, Kaneko S (2002) Matrix metalloproteinase (MMP)-2 and MMP-9 activities in human seminal plasma. Mol Hum Reprod 8: 32-36.
- Shivaprasad HV, Rajesh R, Nanda BL, Dharmappa KK, Vishwanath BS (2009) Thrombin like activity of Asclepias curassavica L. latex: action of cysteine proteases. J Ethnopharmacol 123: 106-109.
- Shyamalamma S, Chandra SB, Hegde M, Naryanswamy P (2008) Evaluation of genetic diversity in jackfruit (Artocarpus heterophyllus Lam.) based on amplified fragment length polymorphism markers. Genet Mol Res 7: 645-656.
- Sreedha AS, Vanathi P, Rao K (2010) Stress proteins in biology and medicine: evolution, adaptation and clinical evaluation. Int J Pharma Bio Scis 1: 1-36.
- Tian M, Huitema E, Da Cunha L, Torto-Alalibo T, Kamoun S (2004) A Kazal-like extracellular serine protease inhibitor from Phytophthora infestans targets the tomato pathogenesis-related protease P69B. J Biol Chem 279: 26370-26377.
- Toma W, Hiruma-Lima CA, Guerrero RO, Brito AR (2005) Preliminary studies of Mammea americana L. (Guttiferae) bark/latex extract point to an effective antiulcer effect on gastric ulcer models in mice. Phytomedicine 12: 345-350.
- Wang W, Vinocur B, Shoseyov O, Altman A (2004) Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. Trends Plant Sci 9: 244-252.