Recognition sequences and structural elements contribute to shedding susceptibility of membrane proteins (original) (raw)

Abstract

Although regulated ectodomain shedding affects a large panel of structurally and functionally unrelated proteins, little is known about the mechanisms controlling this process. Despite a lack of sequence similarities around cleavage sites, most proteins are shed in response to the stimulation of protein kinase C by phorbol esters. The signal-transducing receptor subunit gp130 is not a substrate of the regulated shedding machinery. We generated several chimaeric proteins of gp130 and the proteins tumour necrosis factor alpha (TNF-alpha), transforming growth factor alpha (TGF-alpha) and interleukin 6 receptor (IL-6R), which are known to be subject to shedding. By exchanging small peptide sequences of gp130 for cleavage-site peptides of TNF-alpha, TGF-alpha and IL-6R we showed that these short sequences conferred susceptibility to spontaneous and phorbol-ester-induced shedding of gp130. Importantly, these chimaeric gp130 proteins were functional, as shown by the phosphorylation of gp130 and the activation of signal transduction and activators of transcription 3 ('STAT3') on stimulation with cytokine. To investigate minimal requirements for shedding, truncated cleavage-site peptides of IL-6R were inserted into gp130. The resulting chimaeras were susceptible to shedding and showed the same cleavage pattern as observed in the chimaeras containing the complete IL-6R cleavage site. Surprisingly, we could also generate cleavable chimaeras by exchanging the juxtamembrane sequence of gp130 for the corresponding region of leukaemia inhibitory factor ('LIF') receptor, a protein that like gp130 is not subject to regulated or spontaneous shedding. Thus it seems that there is no minimal consensus shedding sequence. We speculate that structural changes allow the access of the protease to a membrane-proximal region, leading to shedding of the protein.

Full Text

The Full Text of this article is available as a PDF (301.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Althoff K., Reddy P., Voltz N., Rose-John S., Müllberg J. Shedding of interleukin-6 receptor and tumor necrosis factor alpha. Contribution of the stalk sequence to the cleavage pattern of transmembrane proteins. Eur J Biochem. 2000 May;267(9):2624–2631. doi: 10.1046/j.1432-1327.2000.01278.x. [DOI] [PubMed] [Google Scholar]
  2. Arribas J., Coodly L., Vollmer P., Kishimoto T. K., Rose-John S., Massagué J. Diverse cell surface protein ectodomains are shed by a system sensitive to metalloprotease inhibitors. J Biol Chem. 1996 May 10;271(19):11376–11382. doi: 10.1074/jbc.271.19.11376. [DOI] [PubMed] [Google Scholar]
  3. Arribas J., López-Casillas F., Massagué J. Role of the juxtamembrane domains of the transforming growth factor-alpha precursor and the beta-amyloid precursor protein in regulated ectodomain shedding. J Biol Chem. 1997 Jul 4;272(27):17160–17165. doi: 10.1074/jbc.272.27.17160. [DOI] [PubMed] [Google Scholar]
  4. Bazan J. F. Haemopoietic receptors and helical cytokines. Immunol Today. 1990 Oct;11(10):350–354. doi: 10.1016/0167-5699(90)90139-z. [DOI] [PubMed] [Google Scholar]
  5. Blobel C. P. Metalloprotease-disintegrins: links to cell adhesion and cleavage of TNF alpha and Notch. Cell. 1997 Aug 22;90(4):589–592. doi: 10.1016/s0092-8674(00)80519-x. [DOI] [PubMed] [Google Scholar]
  6. Brachmann R., Lindquist P. B., Nagashima M., Kohr W., Lipari T., Napier M., Derynck R. Transmembrane TGF-alpha precursors activate EGF/TGF-alpha receptors. Cell. 1989 Feb 24;56(4):691–700. doi: 10.1016/0092-8674(89)90591-6. [DOI] [PubMed] [Google Scholar]
  7. Brakebusch C., Varfolomeev E. E., Batkin M., Wallach D. Structural requirements for inducible shedding of the p55 tumor necrosis factor receptor. J Biol Chem. 1994 Dec 23;269(51):32488–32496. [PubMed] [Google Scholar]
  8. Bravo J., Staunton D., Heath J. K., Jones E. Y. Crystal structure of a cytokine-binding region of gp130. EMBO J. 1998 Mar 16;17(6):1665–1674. doi: 10.1093/emboj/17.6.1665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Crowe P. D., Walter B. N., Mohler K. M., Otten-Evans C., Black R. A., Ware C. F. A metalloprotease inhibitor blocks shedding of the 80-kD TNF receptor and TNF processing in T lymphocytes. J Exp Med. 1995 Mar 1;181(3):1205–1210. doi: 10.1084/jem.181.3.1205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Diamant M., Rieneck K., Mechti N., Zhang X. G., Svenson M., Bendtzen K., Klein B. Cloning and expression of an alternatively spliced mRNA encoding a soluble form of the human interleukin-6 signal transducer gp130. FEBS Lett. 1997 Jul 28;412(2):379–384. doi: 10.1016/s0014-5793(97)00750-3. [DOI] [PubMed] [Google Scholar]
  11. Ehlers M. R., Schwager S. L., Scholle R. R., Manji G. A., Brandt W. F., Riordan J. F. Proteolytic release of membrane-bound angiotensin-converting enzyme: role of the juxtamembrane stalk sequence. Biochemistry. 1996 Jul 23;35(29):9549–9559. doi: 10.1021/bi9602425. [DOI] [PubMed] [Google Scholar]
  12. Fan H., Derynck R. Ectodomain shedding of TGF-alpha and other transmembrane proteins is induced by receptor tyrosine kinase activation and MAP kinase signaling cascades. EMBO J. 1999 Dec 15;18(24):6962–6972. doi: 10.1093/emboj/18.24.6962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fann M. J., Patterson P. H. Neuropoietic cytokines and activin A differentially regulate the phenotype of cultured sympathetic neurons. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):43–47. doi: 10.1073/pnas.91.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fitzgerald M. L., Wang Z., Park P. W., Murphy G., Bernfield M. Shedding of syndecan-1 and -4 ectodomains is regulated by multiple signaling pathways and mediated by a TIMP-3-sensitive metalloproteinase. J Cell Biol. 2000 Feb 21;148(4):811–824. doi: 10.1083/jcb.148.4.811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fourcin M., Chevalier S., Guillet C., Robledo O., Froger J., Pouplard-Barthelaix A., Gascan H. gp130 transducing receptor cross-linking is sufficient to induce interleukin-6 type responses. J Biol Chem. 1996 May 17;271(20):11756–11760. doi: 10.1074/jbc.271.20.11756. [DOI] [PubMed] [Google Scholar]
  16. Gearing A. J., Beckett P., Christodoulou M., Churchill M., Clements J., Davidson A. H., Drummond A. H., Galloway W. A., Gilbert R., Gordon J. L. Processing of tumour necrosis factor-alpha precursor by metalloproteinases. Nature. 1994 Aug 18;370(6490):555–557. doi: 10.1038/370555a0. [DOI] [PubMed] [Google Scholar]
  17. Gearing D. P., Thut C. J., VandeBos T., Gimpel S. D., Delaney P. B., King J., Price V., Cosman D., Beckmann M. P. Leukemia inhibitory factor receptor is structurally related to the IL-6 signal transducer, gp130. EMBO J. 1991 Oct;10(10):2839–2848. doi: 10.1002/j.1460-2075.1991.tb07833.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Giri J. G., Ahdieh M., Eisenman J., Shanebeck K., Grabstein K., Kumaki S., Namen A., Park L. S., Cosman D., Anderson D. Utilization of the beta and gamma chains of the IL-2 receptor by the novel cytokine IL-15. EMBO J. 1994 Jun 15;13(12):2822–2830. doi: 10.1002/j.1460-2075.1994.tb06576.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hibi M., Murakami M., Saito M., Hirano T., Taga T., Kishimoto T. Molecular cloning and expression of an IL-6 signal transducer, gp130. Cell. 1990 Dec 21;63(6):1149–1157. doi: 10.1016/0092-8674(90)90411-7. [DOI] [PubMed] [Google Scholar]
  20. Hibi M., Nakajima K., Hirano T. IL-6 cytokine family and signal transduction: a model of the cytokine system. J Mol Med (Berl) 1996 Jan;74(1):1–12. doi: 10.1007/BF00202068. [DOI] [PubMed] [Google Scholar]
  21. Hooper N. M., Karran E. H., Turner A. J. Membrane protein secretases. Biochem J. 1997 Jan 15;321(Pt 2):265–279. doi: 10.1042/bj3210265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kallen K. J., Grötzinger J., Lelièvre E., Vollmer P., Aasland D., Renné C., Müllberg J., Myer zum Büschenfelde K. H., Gascan H., Rose-John S. Receptor recognition sites of cytokines are organized as exchangeable modules. Transfer of the leukemia inhibitory factor receptor-binding site from ciliary neurotrophic factor to interleukin-6. J Biol Chem. 1999 Apr 23;274(17):11859–11867. doi: 10.1074/jbc.274.17.11859. [DOI] [PubMed] [Google Scholar]
  23. Kernebeck T., Pflanz S., Müller-Newen G., Kurapkat G., Scheek R. M., Dijkstra K., Heinrich P. C., Wollmer A., Grzesiek S., Grötzinger J. The signal transducer gp130: solution structure of the carboxy-terminal domain of the cytokine receptor homology region. Protein Sci. 1999 Jan;8(1):5–12. doi: 10.1110/ps.8.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kiessling L. L., Gordon E. J. Transforming the cell surface through proteolysis. Chem Biol. 1998 Mar;5(3):R49–R62. doi: 10.1016/s1074-5521(98)90056-4. [DOI] [PubMed] [Google Scholar]
  25. Lammich S., Kojro E., Postina R., Gilbert S., Pfeiffer R., Jasionowski M., Haass C., Fahrenholz F. Constitutive and regulated alpha-secretase cleavage of Alzheimer's amyloid precursor protein by a disintegrin metalloprotease. Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3922–3927. doi: 10.1073/pnas.96.7.3922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Li Y. M., Lai M. T., Xu M., Huang Q., DiMuzio-Mower J., Sardana M. K., Shi X. P., Yin K. C., Shafer J. A., Gardell S. J. Presenilin 1 is linked with gamma-secretase activity in the detergent solubilized state. Proc Natl Acad Sci U S A. 2000 May 23;97(11):6138–6143. doi: 10.1073/pnas.110126897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Massagué J., Pandiella A. Membrane-anchored growth factors. Annu Rev Biochem. 1993;62:515–541. doi: 10.1146/annurev.bi.62.070193.002503. [DOI] [PubMed] [Google Scholar]
  28. McGeehan G. M., Becherer J. D., Bast R. C., Jr, Boyer C. M., Champion B., Connolly K. M., Conway J. G., Furdon P., Karp S., Kidao S. Regulation of tumour necrosis factor-alpha processing by a metalloproteinase inhibitor. Nature. 1994 Aug 18;370(6490):558–561. doi: 10.1038/370558a0. [DOI] [PubMed] [Google Scholar]
  29. McMahan C. J., Slack J. L., Mosley B., Cosman D., Lupton S. D., Brunton L. L., Grubin C. E., Wignall J. M., Jenkins N. A., Brannan C. I. A novel IL-1 receptor, cloned from B cells by mammalian expression, is expressed in many cell types. EMBO J. 1991 Oct;10(10):2821–2832. doi: 10.1002/j.1460-2075.1991.tb07831.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Merlos-Suárez A., Arribas J. Mechanisms controlling the shedding of transmembrane molecules. Biochem Soc Trans. 1999 Feb;27(2):243–246. doi: 10.1042/bst0270243. [DOI] [PubMed] [Google Scholar]
  31. Migaki G. I., Kahn J., Kishimoto T. K. Mutational analysis of the membrane-proximal cleavage site of L-selectin: relaxed sequence specificity surrounding the cleavage site. J Exp Med. 1995 Aug 1;182(2):549–557. doi: 10.1084/jem.182.2.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Mohler K. M., Sleath P. R., Fitzner J. N., Cerretti D. P., Alderson M., Kerwar S. S., Torrance D. S., Otten-Evans C., Greenstreet T., Weerawarna K. Protection against a lethal dose of endotoxin by an inhibitor of tumour necrosis factor processing. Nature. 1994 Jul 21;370(6486):218–220. doi: 10.1038/370218a0. [DOI] [PubMed] [Google Scholar]
  33. Mosley B., De Imus C., Friend D., Boiani N., Thoma B., Park L. S., Cosman D. Dual oncostatin M (OSM) receptors. Cloning and characterization of an alternative signaling subunit conferring OSM-specific receptor activation. J Biol Chem. 1996 Dec 20;271(51):32635–32643. doi: 10.1074/jbc.271.51.32635. [DOI] [PubMed] [Google Scholar]
  34. Müllberg J., Althoff K., Jostock T., Rose-John S. The importance of shedding of membrane proteins for cytokine biology. Eur Cytokine Netw. 2000 Mar;11(1):27–38. [PubMed] [Google Scholar]
  35. Müllberg J., Dittrich E., Graeve L., Gerhartz C., Yasukawa K., Taga T., Kishimoto T., Heinrich P. C., Rose-John S. Differential shedding of the two subunits of the interleukin-6 receptor. FEBS Lett. 1993 Oct 11;332(1-2):174–178. doi: 10.1016/0014-5793(93)80507-q. [DOI] [PubMed] [Google Scholar]
  36. Müllberg J., Oberthür W., Lottspeich F., Mehl E., Dittrich E., Graeve L., Heinrich P. C., Rose-John S. The soluble human IL-6 receptor. Mutational characterization of the proteolytic cleavage site. J Immunol. 1994 May 15;152(10):4958–4968. [PubMed] [Google Scholar]
  37. Müllberg J., Rauch C. T., Wolfson M. F., Castner B., Fitzner J. N., Otten-Evans C., Mohler K. M., Cosman D., Black R. A. Further evidence for a common mechanism for shedding of cell surface proteins. FEBS Lett. 1997 Jan 20;401(2-3):235–238. doi: 10.1016/s0014-5793(96)01480-9. [DOI] [PubMed] [Google Scholar]
  38. Müllberg J., Schooltink H., Stoyan T., Günther M., Graeve L., Buse G., Mackiewicz A., Heinrich P. C., Rose-John S. The soluble interleukin-6 receptor is generated by shedding. Eur J Immunol. 1993 Feb;23(2):473–480. doi: 10.1002/eji.1830230226. [DOI] [PubMed] [Google Scholar]
  39. Müllberg J., Schooltink H., Stoyan T., Heinrich P. C., Rose-John S. Protein kinase C activity is rate limiting for shedding of the interleukin-6 receptor. Biochem Biophys Res Commun. 1992 Dec 15;189(2):794–800. doi: 10.1016/0006-291x(92)92272-y. [DOI] [PubMed] [Google Scholar]
  40. Renné C., Kallen K. J., Müllberg J., Jostock T., Grötzinger J., Rose-John S. A new type of cytokine receptor antagonist directly targeting gp130. J Biol Chem. 1998 Oct 16;273(42):27213–27219. doi: 10.1074/jbc.273.42.27213. [DOI] [PubMed] [Google Scholar]
  41. Sadhukhan R., Santhamma K. R., Reddy P., Peschon J. J., Black R. A., Sen I. Unaltered cleavage and secretion of angiotensin-converting enzyme in tumor necrosis factor-alpha-converting enzyme-deficient mice. J Biol Chem. 1999 Apr 9;274(15):10511–10516. doi: 10.1074/jbc.274.15.10511. [DOI] [PubMed] [Google Scholar]
  42. Schlöndorff J., Becherer J. D., Blobel C. P. Intracellular maturation and localization of the tumour necrosis factor alpha convertase (TACE). Biochem J. 2000 Apr 1;347(Pt 1):131–138. [PMC free article] [PubMed] [Google Scholar]
  43. Schwager S. L., Chubb A. J., Scholle R. R., Brandt W. F., Eckerskorn C., Sturrock E. D., Ehlers M. R. Phorbol ester-induced juxtamembrane cleavage of angiotensin-converting enzyme is not inhibited by a stalk containing intrachain disulfides. Biochemistry. 1998 Nov 3;37(44):15449–15456. doi: 10.1021/bi981260k. [DOI] [PubMed] [Google Scholar]
  44. Senaldi G., Varnum B. C., Sarmiento U., Starnes C., Lile J., Scully S., Guo J., Elliott G., McNinch J., Shaklee C. L. Novel neurotrophin-1/B cell-stimulating factor-3: a cytokine of the IL-6 family. Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11458–11463. doi: 10.1073/pnas.96.20.11458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Sisodia S. S. Beta-amyloid precursor protein cleavage by a membrane-bound protease. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6075–6079. doi: 10.1073/pnas.89.13.6075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Taga T., Kishimoto T. Gp130 and the interleukin-6 family of cytokines. Annu Rev Immunol. 1997;15:797–819. doi: 10.1146/annurev.immunol.15.1.797. [DOI] [PubMed] [Google Scholar]
  47. Tang P., Hung M-C, Klostergaard J. Length of the linking domain of human pro-tumor necrosis factor determines the cleavage processing. Biochemistry. 1996 Jun 25;35(25):8226–8233. doi: 10.1021/bi952183l. [DOI] [PubMed] [Google Scholar]
  48. Vassar R., Bennett B. D., Babu-Khan S., Kahn S., Mendiaz E. A., Denis P., Teplow D. B., Ross S., Amarante P., Loeloff R. Beta-secretase cleavage of Alzheimer's amyloid precursor protein by the transmembrane aspartic protease BACE. Science. 1999 Oct 22;286(5440):735–741. doi: 10.1126/science.286.5440.735. [DOI] [PubMed] [Google Scholar]
  49. Vollmer P., Walev I., Rose-John S., Bhakdi S. Novel pathogenic mechanism of microbial metalloproteinases: liberation of membrane-anchored molecules in biologically active form exemplified by studies with the human interleukin-6 receptor. Infect Immun. 1996 Sep;64(9):3646–3651. doi: 10.1128/iai.64.9.3646-3651.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Vriend G. Parameter relation rows: a query system for protein structure function relationships. Protein Eng. 1990 Dec;4(2):221–223. doi: 10.1093/protein/4.2.221. [DOI] [PubMed] [Google Scholar]
  51. Walev I., Vollmer P., Palmer M., Bhakdi S., Rose-John S. Pore-forming toxins trigger shedding of receptors for interleukin 6 and lipopolysaccharide. Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):7882–7887. doi: 10.1073/pnas.93.15.7882. [DOI] [PMC free article] [PubMed] [Google Scholar]