Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
권호 표지
DOI QR코드

DOI QR Code

Carboxypeptidase E, Identified As a Direct Interactor of Growth Hormone, Is Important for Efficient Secretion of the Hormone

  • Mizutani, Akiko (Basic Medical Science and Molecular Medicine, Tokai University School of Medicine) ;
  • Inoko, Hidetoshi (Basic Medical Science and Molecular Medicine, Tokai University School of Medicine) ;
  • Tanaka, Masafumi (Basic Medical Science and Molecular Medicine, Tokai University School of Medicine)
  • Received : 2016.07.20
  • Accepted : 2016.09.22
  • Published : 2016.10.31
Download PDF
⟨ Previous Next ⟩

Abstract

We have identified 88 interactor candidates for human growth hormone (GH) by the yeast two-hybrid assay. Among those, we focused our efforts on carboxypeptidase E (CPE), which has been thought to play a key role in sorting prohormones, such as pro-opiomelanocortin (POMC), to regulated secretory vesicles. We found that CPE colocalizes with and interacts with GH in AtT20 pituitary cells. Downregulation of CPE led to decreased levels of GH secretion, consistent with involvement of CPE in GH sorting/secretion. Our binding assay in vitro with bacterially expressed proteins suggested that GH directly interacts with CPE but in a manner different from POMC.

Keywords

References

  1. Ballinger, A., Camacho-Hubner, C., and Croft, N. (2001). Growth failure and intestinal inflammation. Qjm 94, 121-125. https://doi.org/10.1093/qjmed/94.3.121
  2. Burgess, T.L., and Kelly, R.B. (1987). Constitutive and regulated secretion of proteins. Annu. Rev. Cell Biol. 3, 243-293. https://doi.org/10.1146/annurev.cb.03.110187.001331
  3. Cawley, N.X., Rodriguez, Y.M., Maldonado, A., and Loh, Y.P. (2003). Trafficking of mutant carboxypeptidase E to secretory granules in a beta-cell line derived from Cpe(fat)/Cpe(fat) mice. Endocrinology 144, 292-298. https://doi.org/10.1210/en.2002-220588
  4. Cawley, N.X., Wetsel, W.C., Murthy, S.R., Park, J.J., Pacak, K., and Loh, Y.P. (2012). New roles of carboxypeptidase E in endocrine and neural function and cancer. Endocr. Rev. 33, 216-253. https://doi.org/10.1210/er.2011-1039
  5. Cool, D.R., and Loh, Y.P. (1998). Carboxypeptidase E is a sorting receptor for prohormones: binding and kinetic studies. Mol. Cell. Endocrinol. 139, 7-13. https://doi.org/10.1016/S0303-7207(98)00081-1
  6. Cool, D.R., Fenger, M., Snell, C.R., and Loh, Y.P. (1995). Identification of the sorting signal motif within pro-opiomelanocortin for the regulated secretory pathway. J. Biol. Chem. 270, 8723-8729. https://doi.org/10.1074/jbc.270.15.8723
  7. Cool, D.R., Normant, E., Shen, F.-S., Chen, H.-C., Pannell, L., Zhang, Y., and Loh, Y.P. (1997). Carboxypeptidase E is a regulated secretory pathway sorting receptor: genetic obliteration leads to endocrine disorders in $Cpe^{fat}$ mice. Cell 88, 73-83. https://doi.org/10.1016/S0092-8674(00)81860-7
  8. Dannies, P.S. (2002). Mechanisms for storage of prolactin and growth hormone in secretory granules. Mol. Genet. Metab. 76, 6-13. https://doi.org/10.1016/S1096-7192(02)00011-2
  9. Davidson, H.W., and Hutton, J.C. (1987). The insulin-secretory-granule carboxypeptidase H. Purification and demonstration of involvement in proinsulin processing. Biochem. J. 245, 575-582. https://doi.org/10.1042/bj2450575
  10. Ewing, R.M., Chu, P., Elisma, F., Li, H., Taylor, P., Climie, S., McBroom-Cerajewski, L., Robinson, M.D., O'Connor, L., and Li, M. (2007). Large-scale mapping of human protein-protein interactions by mass spectrometry. Mol. Sys. Biol. 3, 89.
  11. Hales, C.M., Griner, R., Hobdy-Henderson, K.C., Dorn, M.C., Hardy, D., Kumar, R., Navarre, J., Chan, E.K., Lapierre, L.A., and Goldenring, J.R. (2001). Identification and characterization of a family of Rab11-interacting proteins. J. Biol. Chem. 276, 39067-39075. https://doi.org/10.1074/jbc.M104831200
  12. Hook, V., and Loh, Y.P. (1984). Carboxypeptidase B-like converting enzyme activity in secretory granules of rat pituitary. Proc. Natl. Acad. Sci. USA 81, 2776-2780. https://doi.org/10.1073/pnas.81.9.2776
  13. Hosaka, M., Watanabe, T., Sakai, Y., Kato, T., and Takeuchi, T. (2005). Interaction between secretogranin III and carboxypeptidase E facilitates prohormone sorting within secretory granules. J. Cell Sci. 118, 4785-4795. https://doi.org/10.1242/jcs.02608
  14. Irminger, J.-C., Verchere, C.B., Meyer, K., and Halban, P.A. (1997). Proinsulin Targeting to the regulated pathway is not impaired in carboxypeptidase E-deficient $Cpe^{fat}/Cpe^{fat}$ mice. J. Biol. Chem. 272, 27532-27534. https://doi.org/10.1074/jbc.272.44.27532
  15. Kemppainen, R.J., and Behrend, E.N. (2010). Acute inhibition of carboxypeptidase E expression in AtT-20 cells does not affect regulated secretion of ACTH. Regul. Pept. 165, 174-179. https://doi.org/10.1016/j.regpep.2010.07.162
  16. Khvotchev, M.V., Ren, M., Takamori, S., Jahn, R., and Sudhof, T.C. (2003). Divergent functions of neuronal Rab11b in $Ca^{2+}$-regulated versus constitutive exocytosis. J. Neurosci. 23, 10531-10539. https://doi.org/10.1523/JNEUROSCI.23-33-10531.2003
  17. Lou, H., Kim, S.K., Zaitsev, E., Snell, C.R., Lu, B., and Loh, Y.P. (2005). Sorting and activity-dependent secretion of BDNF require interaction of a specific motif with the sorting receptor carboxypeptidase E. Neuron 45, 245-255. https://doi.org/10.1016/j.neuron.2004.12.037
  18. McGirr, R., Guizzetti, L., and Dhanvantari, S. (2013). The sorting of proglucagon to secretory granules is mediated by carboxypeptidase E and intrinsic sorting signals. J. Endocrinol. 217, 229-240. https://doi.org/10.1530/JOE-12-0468
  19. Moore, H.P., Andresen, J.M., Eaton, B.A., Grabe, M., Haugwitz, M., Wu, M.M., and Machen, T.E. (2002). Biosynthesis and secretion of pituitary hormones: dynamics and regulation. Arch. Physiol. Biochem. 110, 16-25. https://doi.org/10.1076/apab.110.1.16.903
  20. Normant, E., and Loh, Y.P. (1998). Depletion of carboxypeptidase E, a regulated secretory pathway sorting receptor, causes misrouting and constitutive secretion of proinsulin and proenkephalin, but not chromogranin A. Endocrinology 139, 2137-2145. https://doi.org/10.1210/endo.139.4.5951
  21. Park, J.J., Cawley, N.X., and Loh, Y.P. (2008). Carboxypeptidase E cytoplasmic tail-driven vesicle transport is key for activity-dependent secretion of peptide hormones. Mol. Endocrinol. 22, 989-1005. https://doi.org/10.1210/me.2007-0473
  22. Rivas, R.J., and Moore, H. (1989). Spatial segregation of the regulated and constitutive secretory pathways. J. Cell Biol. 109, 51-60. https://doi.org/10.1083/jcb.109.1.51
  23. Shaner, N.C., Lin, M.Z., McKeown, M.R., Steinbach, P.A., Hazelwood, K.L., Davidson, M.W., and Tsien, R.Y. (2008). Improving the photostability of bright monomeric orange and red fluorescent proteins. Nat. Methods 5, 545-551. https://doi.org/10.1038/nmeth.1209
  24. Shen, F.-S., and Loh, Y.P. (1997). Intracellular misrouting and abnormal secretion of adrenocorticotropin and growth hormone in $Cpe^{fat}$ mice associated with a carboxypeptidase E mutation. Proc. Natl. Acad. Sci. USA 94, 5314-5319. https://doi.org/10.1073/pnas.94.10.5314
  25. Shu, X., Cheng, Y., and Edwards, R.H. (2013). Self-assembly of VPS41 promotes sorting required for biogenesis of the regulated secretory pathway. Dev. Cell 27, 1-13. https://doi.org/10.1016/j.devcel.2013.09.027
  26. Tanaka, M. (1996). Modulation of promoter occupancy by cooperative DNA binding and activation-domain function is a major determinant of transcriptional regulation by activators in vivo. Proc. Natl. Acad. Sci. USA 93, 4311-4315. https://doi.org/10.1073/pnas.93.9.4311
  27. Tanaka, M., and Herr, W. (1990). Differential transcriptional activation by Oct-1 and Oct-2: interdependent activation domains induce Oct-2 phosphorylation. Cell 60, 375-386. https://doi.org/10.1016/0092-8674(90)90589-7
  28. Wang, J., Cawley, N.X., Voutetakis, A., Rodriguez, Y.M., Goldsmith, C.M., Nieman, L.K., Hoque, A.S., Frank, S.J., Snell, C.R., and Loh, Y.P. (2005). Partial redirection of transgenic human growth hormone secretion from rat salivary glands. Hum. Gene Ther. 16, 571-583. https://doi.org/10.1089/hum.2005.16.571
  29. Zhang, C.-F., Snell, C.R., and Peng Loh, Y. (1999). Identification of a novel prohormone sorting signal-binding site on carboxypeptidase E, a regulated secretory pathway-sorting receptor. Mol. Endocrinol. 13, 527-536. https://doi.org/10.1210/mend.13.4.0267

Cited by

  1. Metal-dependent hormone function: the emerging interdisciplinary field of metalloendocrinology pp.1756-591X, 2019, https://doi.org/10.1039/C8MT00221E
  2. Carboxypeptidase E down-regulation regulates transcriptional and epigenetic profiles in pancreatic cancer cell line: A network analysis vol.29, pp.1, 2016, https://doi.org/10.3233/cbm-191163