purification of hyaluronan binding proteins from human normal and cancer serum

background: analysis of human cancer serum has revealed the presence of high amounts of hyaluronan (hyaluronic acid, ha) when compared to human normal serum, it is well documented that ha and its receptors, known as hyaladherins (habps) are involved in matrix regulation, cell proliferation, migration and malignant tumour progression. these hyaladherins not only interact with hyaluronan at the matrix proper but also with  hyaluronan at the plasma membrane as a cell surface receptors and thus influence cell physiology including secretion of this protein into the circulatory system. methods: normal serum and colon cancer serum samples were included in this study, using biochemical techniques such as gel permeation, strong anion exchange chromatography, single dimension electrophoresis and western blot analysis. results: this study is based on the clinical work of normal serum and colon cancer serum. the description of the procedure was given for the fractionation of serum proteins mainly habps from 20 normal and 15 colon cancer patients by using special biotinylated hyaluronan probe. conclusion: to evaluate whether serum habps levels could be used as diagnostic marker for human cancer. the semi purified serum from normal and colon cancer patients showed mainly a major protein (57kda) and a minor one (30kda) by overlay experiments with b-ha probe and these results were confirmed by competition experiments with cold ha. please cite this article in: fekry  b, siddaiah m, srinivas kp, nunna rv, banerjee sd, mortha kk purification of hyaluronan binding proteins from human normal and cancer serum. iran j cancer prev, 2010; vol3, no2, p.79-86. key words: hyaluronan (hyaluronic acid, ha) hyaluronic acid binding protein (habp); biotinylated hyaluronan probe (bha); cold ha. references 1. knudson w, biswas c, li xq, nemec re, toole bp. the role and regulation of tumor-associated hyaluronan. ciba found symp. 1989; 143: 150-9. 2. toole bp. hyaluronan and its binding proteins, the hyaladherins. curr opin cell biol. 1990; 2: 839-44. 3. laurent tc, fraser jre. hyaluronan. fasebj; 1992, 6: 2397-2404. 4. turley ea, torrance j. localization of hyaluronate and hyaluronate-binding protein on motile and non-motile fibroblasts. exp cell res. 1985; 161: 17-28. 5. toole bp. glycosaminoglycans in morphogenesis. in, cell biology of extracellular matrix.1981, hay ed, ed. new york: plenum press; 259-94. 6. laurent tc, fraser jre. the properties and turnover of hyaluronan ciba found symp. 1986; 124:9-29. 7. laurent tc, dahl ims, dahl lb ,engstrom laurent a, eriksson s, fraser jre, et al. the catabolic fate of hyaluronic acid. connect tissue res; 1986, 15:33-41. 8. engstrom laurent a, loof, l, nyberg a, schroder t. increased serum levels of hyaluronate in liver disease. hepatology .1985; 5:638-42. 9. frebourg t, delpech b, bercoff e. serum hyaluronate in liver diseases: study by enzymoimmunological assay. hepatology.1986,6: 392-95. 10. inger  s , laurent t.c. concentration of hyaluronan in the serum of untreated cancerpatients with special reference to patients with mesothelioma.cancer.1988, 62:326-330. 11. marhaba r, zoller m. cd44 in cancer progression: adhesion, migration and growth regulation.j. mol  histol. 2004; 35: 211-31. 12. toole b.  proteoglycans and hyaluronan in morphogenesis and differentiation. in: hay e, editor. cell biology of extracellular matrix. 2nd edition. new york: plenum press; 1991:. 305–342. 13. entwistle j, hall c.l, turley e.a.  ha receptors: regulators of signalling to the  cytoskeleton.j.cell.biochem. 1998;61: 569-577. 14. auvinen p.k, parkkinen j.j, johansson r.t, agren u.m, tammi r.h, eskelinen m.j, kosma v.m. expression of hyaluronan in benign and malignant breast lesions. int j cancer. 1997; 74: 477-81. 15. sy m.s, guo y.j and stamenkovic i. distinct effects of two cd44 isoforms on tumor growth in vivo.j exp med. 1991; 174: 859-66. 16. takahashi k, stamenkovic i, cutler m, saya h, tanabe k.k.  cd44 hyaluronate binding influences growth kinetics and tumorigenicity of human colon carcinoma.oncogene. 1995; 11: 2223-32. 17. rhodes j m. usefulness of novel tumour markers annals of oncology. 1999; 10: s118-s121. 18. jacobs i , bast rc jr.the ca 125 tumour-associated antigen.hum reprod. 1989; 4(1):1-12. 19. toole bp: hyaluronan, in proteoglycans; structure, biology and molecular interactions 2000, edited by: iozzo rv. marcel dekker, new york, 61-92. 20. laurent tc, laurent ub, fraser jr: the structure and function ofhyaluronan: an overview. immunol cell biol. 1996; 74:a1-a7. 21. collis l, hall c, lange l, ziebel m, prestwich r, turley ea. rapidhyaluronan uptake is associated with enhanced motility:implication for an intracellular mode of action. febs lett. 1998; 440:444-9. 22. wight t.n, merrilees m.j. proteoglycans in atherosclerosis and restenosis: key roles for versican.circ. res. 2004; 94:1158–67