Skip to main content
SpringerLink
Log in

Glycosylation and over-expression of endometriosis-associated peritoneal haptoglobin

  • Published:
Glycoconjugate Journal Aims and scope Submit manuscript

Abstract

Peritoneal endometriotic tissues synthesize and secrete haptoglobin (pHp), which has an analogous nucleotide sequence to hepatic haptoglobin found in serum (sHp). This study performed enzymatic digestions and lectin binding assays to determine if differences in protein glycosylation exist between sHp and pHp, which may provide insight into pHp function and/or identify epitopes for development of novel methods of medical management of endometriosis. To reduce the dependence on surgical collection of peritoneal tissues from women, recombinant peritoneal Hp (rpHp) was produced and its glycosylation analyzed for future functional studies. These results showed the apparent molecular weight of pHp was 3 kDa smaller than sHp. Desialylation and complete N-deglycosylation elicited similar shifts in sHp and pHp electrophoretic migration, suggesting similar sialic acid content and indicating the 3 kDa variance was due to carbohydrate content, not protein degradation, respectively. Sequential deglycosylation of the four sHp N-glycan chains caused a 3 kDa shift per N-glycan removed suggesting the 3 kDa difference between sHp and pHp may be one N-glycan chain. Lectin ELISA and lectin-blotting analyses demonstrated increased pHp and rpHp interactions with MAL and LTL but no difference in binding to SNL compared to sHp from healthy individuals, identifying variations in the ratios of α(2-3) to α(2-6) sialic acid and fucose residues. Recombinant pHp was 100-fold over-expressed with a similar glycosylation pattern to pHp, albeit in an unprocessed α-β Hp polypeptide form. These results are the first to identify differences between pHp and sHp glycosylation and lay groundwork further studies to characterize anomalies in glycan composition and structure, which likely impart pHp with known immunomodulatory functions and may be used as epitopes for development of immune based therapeutics for novel, non-surgical management of endometriosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Sampson JA, Peritoneal endometriosis due to menstrual dissemination of endometrial tissue into the peritoneal cavity, Am J Obstet Gynecol 14, 422–69 (1927).

    Google Scholar 

  2. Nissole M, Donnez J, Peritoneal endometriosis, ovarian endometriosis, and adenomyotic nodules of the rectovaginal septum are three different entities, Fertil Steril 68, 585–98 (1997).

    PubMed  Google Scholar 

  3. Cirkel U, Ochs H, Mues B, Zwadlo G, Sorg C, Schneider HP, Inflammatory reaction in endometriotic tissue: An immunohistochemical study, Eur J Obstet Gynecol Reprod Biol 48, 43–50 (1993).

    PubMed  Google Scholar 

  4. Brosens IA, Endometriosis. Current issues in diagnosis and medical management, J Reprod Med 43, 281–6 (1998).

    PubMed  Google Scholar 

  5. Sharpe KL, Zimmer RL, Griffin WT, Penney LL, Polypeptides synthesized and released by human endometriosis differ from those of the uterine endometrium in cell and tissue explant culture, Fertil Steril 60, 839–51 (1993).

    PubMed  Google Scholar 

  6. Sharpe-Timms KL, Piva M, Ricke EA, Surewicz K, Zhang YL, Zimmer RL, Endometriotic lesions synthesize and secrete a haptoglobin-like protein, Biol Reprod 58, 988–94 (1998).

    PubMed  Google Scholar 

  7. Piva M, Sharpe-Timms KL, Peritoneal endometriotic lesions differentially express a haptoglobin-like gene, Mol Hum Reprod 5, 71–8 (1999).

    PubMed  Google Scholar 

  8. Piva M, Horowitz GM, Sharpe-Timms KL, Interleukin-6 differentially stimulates haptoglobin production by peritoneal and endometriotic cells in vitro: A model for endometrial-peritoneal interaction in endometriosis, J Clin Endocrinol Metab 86, 2553–61 (2001).

    Article  PubMed  Google Scholar 

  9. Sharpe-Timms KL, Ricke EA, Piva M, Horowitz GM, Differential in vivo expression and localization of endometriosis protein-I (ENDO-I), a haptoglobin homologue, in endometrium and endometriotic lesions, Hum Reprod 15, 2180–5 (2000).

    PubMed  Google Scholar 

  10. Sharpe-Timms KL, Zimmer RL, Ricke EA, Piva MA, Horowitz GM, Endometriotic haptoglobin binds peritoneal macrophages and alters their function in endometriosis, Fertil Steril 78, 810–9 (2002).

    PubMed  Google Scholar 

  11. Turner GA, Haptoglobin, A potential reporter molecule for glycosylation changes in disease, Adv Exp Med Biol 376, 231–8 (1995).

    PubMed  Google Scholar 

  12. Nilsson B, Lowe M, Osada J, Ashwell G, Zopf D, The carbohydrate structure of human haptoglobin 1-1. In Glycoconjugates Proceedings of the 6th International Symposium on Glycoconjugates, edited by T Yamakawa, T Osawa, S Handa (Japan Scientific Societies Press, Tokyo, 1981), pp. 275–6.

    Google Scholar 

  13. Katnik I, Jadach J, Krotkiewski H, Gerber J, Investigating the glycosylation of normal and ovarian cancer haptoglobins using digoxigenin-labeled lectins, Glycosyl Dis 1, 97–104 (1994).

    Google Scholar 

  14. Baseler MW, Burrell R, Acute phase reactants in experimental inhalation lung disease, Proc Soc Exp Biol 111, 49–55 (1981).

    Google Scholar 

  15. Baseler MW, Burrell R, Purification of haptoglobin and its effects on lymphocyte and alveolar macrophage responses, Inflamm 7, 387–400 (1983).

    Google Scholar 

  16. Oh SK, Kim SH, Walker JE, Interference with immune response at the level of generating effector cells by tumor-associated haptoglobin, J Natl Canc Inst 82, 934–40 (1990).

    Google Scholar 

  17. Oh SK, Very DL, Walker J, Raam S, Ju ST, An analogy between fetal haptoglobin and a potent immunosuppressant in cancer, Cancer Res 47, 5120–6 (1987).

    PubMed  Google Scholar 

  18. El Ghmati SM, Van Hoeyveld EM, Van Strijp AG, Ceuppens JL, Stevens EAM, Identification of haptoglobin as an alternative ligand for CD11b/CD18, J Immunol 156, 2542–52 (1996).

    PubMed  Google Scholar 

  19. van Dijk W, Turner GA, Mackiewicz A, Changes in glycosylation of acute-phase proteins in health and disease: Occurrence, regulation and function, Glycosyl Dis 1, 5–14 (1994).

    Google Scholar 

  20. van Dijk W, Mackiewicz A, Interleukin-6-type cytokine-induced changes in acute phase protein glycosylation, Ann NY Acad Sci 762, 319–30 (1995).

    PubMed  Google Scholar 

  21. Sharpe-Timms KL, Piva M, Ricke EA, Extrahepatic vs. hepatic haptoglobin in women with endometriosis, Soc. Gynecol. Invest. Chicago, IL Abstract 825 (2000).

    Google Scholar 

  22. Tseng JF, Ryan IP, Milam TD, Murai JT, Schriock ED, Landers DV, Taylor RN, Interleukin-6 secretion in vitro is up-regulated in ectopic and eutopic endometrial stromal cells from women with endometriosis, J Clin Endocrinol Metab 81, 1118–22 (1996).

    PubMed  Google Scholar 

  23. Dobryszycka W, Biological functions of haptoglobin-new pieces to an old puzzle, Euro J Clin Chem Clin Biochem 35, 647–54 (1997).

    Google Scholar 

  24. Langlois MR, Delanghe JR, Biological and clinical significance of haptoglobin polymorphism in humans, Clin Chem 42, 1589–1600 (1996).

    PubMed  Google Scholar 

  25. Thorton BP, Vetvicka V, Pitman M, Goldman RC, Ross GD, Analysis of the sugar specificity and molecular location of the beta-glucan binding lectin site of complement receptor type 3 (CD11b/CD18), J Immunol 156, 1235–46 (1996).

    PubMed  Google Scholar 

  26. Halme J, Becker S, Haskill S, Altered maturation and function of peritoneal macrophages: Possible pathogenesis of endometriosis, Am J Obstet Gynecol 156, 783–9 (1987).

    PubMed  Google Scholar 

  27. van der Straten A, Herzog A, Cabezon T, Bollen A, Characterization of human haptoglobin cDNAs coding for alpha 2FS beta and alpha 1S beta variants, FEBS Lett 168, 103–7 (1984).

    PubMed  Google Scholar 

  28. Piva M, Horowitz G, Sharpe-Timms K, Human peritoneal endometriotic lesions express all three haptoglobin α-chain phenotypes, Biol Reprod 60(Suppl. 1), 261 (1999).

    Google Scholar 

  29. Brinkman-van der Linden CM, de Haan PF, Havenaar EC, van Dijk W, Inflammation-induced expression of sialyl Lewis-X is not restricted to alpha1-acid glycoprotein but also occurs to a lesser extent on alpha1-antichymotrypsin and haptoglobin, Glycoconj J 15, 177–82 (1998).

    PubMed  Google Scholar 

  30. Knibbs RN, Goldstein IJ, Ratcliffe RM, Shibuya N, Characterization of the carbohydrate binding specificity of the leukoagglutinating lectin from Maackia amurensis, J Biol Chem 266, 83–8 (1991).

    PubMed  Google Scholar 

  31. Goodarzi MT, Turner GA, A lectin method for investigating the glycosylation of nanogram amounts of purified glycoprotein, Glycoconj J 14, 493–6 (1997).

    PubMed  Google Scholar 

  32. Yan L, Wilkins PP, Alvarez-Manilla G, Do SI, Smith DF, Cummings RD, Immobilized Lotus tetragonolobus agglutinin binds oligosaccharides containing the Le(x) determinant, Glycoconj J 14, 45–55 (1997).

    PubMed  Google Scholar 

  33. Debray H, Decout D, Strecker G, Spik G, Montreuil J, Specificity of twelve lectins towards oligosaccharides and glycopeptides related to N-Glycosyl proteins, Eur J Biochem 117, 41–55 (1981).

    PubMed  Google Scholar 

  34. Osteen KG, Hill GA, Hargrove JT, Gorstein F, Development of a method to isolate and culture highly purified populations of stromal and epithelial cells from human endometrial biopsy specimens, Fertil Steril 52, 965–72 (1989).

    PubMed  Google Scholar 

  35. Maeda N, DNA polymorphisms in the controlling region of the human haptoglobin genes: A molecular explanation for the haptoglobin 2-1 modified phenotype, Am J Hum Genet 49, 158–66 (1991).

    PubMed  Google Scholar 

  36. Ferens-Sieczkowska M, Midro A, Mierzejewska-Iwanowska B, Zwierz K, Katnik-Prastowska I, Haptoglobin glycoforms in a case of carbohydrate-deficient glycoprotein syndrome, Glycoconj J 16, 573–7 (1999).

    PubMed  Google Scholar 

  37. Ferens-Sieczkowska M, Olczak M, Carbohydrate structures of haptoglobin in sera of healthy people and a patient with congenital disorder of glycosylation, Z Naturforsch 56, 122–31 (2001).

    Google Scholar 

  38. Goodarzi MT, Turner GA, Reproducible and sensitive determination of charged oligosaccharides from haptoglobin by PNGase F digestion and HPAEC/PAD analysis: Glycan composition varies with disease, Glycoconj J 15, 469–75 (1998).

    PubMed  Google Scholar 

  39. Nemansky M, Schiphorst WECM, Van der Eijnden DH, Branching and elongation with lactosaminoglycan chains of N-linked oligosaccharides result in a shift toward termination with α2→3-linked rather than α2→6-linked sialic acid residues, FEBS Lett 363, 280–4 (1995).

    PubMed  Google Scholar 

  40. Turner GA, Goodarzi MT, Thompson S, Glycosylation of alpha-1-proteinase inhibitor and haptoglobin in ovarian cancer: Evidence for two different mechanisms, Glycoconj J 12, 211–8 (1995).

    PubMed  Google Scholar 

  41. Thompson S, Cantwell BM, Matta KL, Turner GA, Parallel changes in the blood levels of abnormally-fucosylated haptoglobin and alpha 1,3 fucosyltransferase in relationship to tumour burden: More evidence for a disturbance of fucose metabolism in cancer, Cancer Lett 65, 115–21 (1992).

    PubMed  Google Scholar 

  42. de Graaf TW, Van der Stelt ME, Anbergen MG, van Dijk W, Inflammation-induced expression of sialyl Lewis X-containing glycan structures on α1-acid glycoprotein (orosomucoid) in human sera, J Exp Med 177, 657–66 (1993).

    PubMed  Google Scholar 

  43. Mann AC, Record CO, Self CH, Turner GA, Monosaccharide composition of haptoglobin in liver diseases and alcohol abuse: Large changes in glycosylation associated with alcoholic liver disease, Clin Chim Acta 227, 69–78 (1994).

    PubMed  Google Scholar 

  44. Dunselman GA, Bouckaert PX, Evers JL, The acute phase response in endometriosis of women, J Reprod Fertil 83, 803–8 (1988).

    PubMed  Google Scholar 

  45. Hanley JM, Haugen TH, Heath EC, Biosynthesis and processing of rat haptoglobin, J Biol Chem 258, 7858–69 (1983).

    PubMed  Google Scholar 

  46. Montreuil J, Recombinant glycoproteins: Pitfalls and strategy. In Polysaccharides in Medical Applications, edited by S Dumitriu (Marcel Dekker, New York, 1996), pp. 481–90.

    Google Scholar 

  47. Raju TS, Stanley P, Gain-of-function Chinese hamster ovary mutants LEC18 and LEC14 each express a novel Nacetylglucosaminyltransferase activity, J Biol Chem 273, 14090–8 (1998).

    PubMed  Google Scholar 

  48. Rothlein R, Springer TA, Complement receptor type threedependent degradation of opsonized erythrocytes by mouse macrophages, J Immunol 135, 2668–72 (1985).

    PubMed  Google Scholar 

  49. Ding A, Wright SW, Nathan C, Activation of murine macrophages by monoclonal antibodies to Mac-1 (complement receptor type 3), J Exper Med 165, 733–49 (1987).

    Google Scholar 

  50. Oh SK, Ross S, Walker JM, Zeisel S, Role of a SER immune suppressor in immune surveillance, Immunology 64, 73–9 (1988).

    PubMed  Google Scholar 

  51. Yong K, Khwaja A, Leukocyte cellular adhesion molecules, Blood Reviews 4, 211–25 (1990).

    PubMed  Google Scholar 

  52. Fan ST, Edgington TS, Integrin regulation of leukocyte inflammatory functions, J Immunol 150, 2972–80 (1993).

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, The University of Missouri, Columbia, MO, 65212, USA

    Marta Piva & Kathy L. Sharpe-Timms

  2. Department of Molecular Biology, Celltech R&D, Inc, Bothell, WA, 98021, USA

    J. Ignacio Moreno

Authors
  1. Marta Piva
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Ignacio Moreno
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kathy L. Sharpe-Timms
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kathy L. Sharpe-Timms.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Piva, M., Moreno, J.I. & Sharpe-Timms, K.L. Glycosylation and over-expression of endometriosis-associated peritoneal haptoglobin. Glycoconj J 19, 33–41 (2002). https://doi.org/10.1023/A:1022580813870

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1022580813870

Search

Navigation