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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1995 Mar 14;92(6):1866–1870. doi: 10.1073/pnas.92.6.1866

Molecular cloning of a ligand for the EPH-related receptor protein-tyrosine kinase Htk.

B D Bennett 1, F C Zeigler 1, Q Gu 1, B Fendly 1, A D Goddard 1, N Gillett 1, W Matthews 1
PMCID: PMC42383  PMID: 7534404

Abstract

Htk is a receptor protein-tyrosine kinase that is related to the EPH subfamily of tyrosine kinases. The receptor has a wide tissue distribution including expression in several myeloid hematopoietic cell lines. Using an Htk-Fc fusion protein, a protein ligand for this receptor was expression cloned from the murine kidney mesangial cell line SV40MES 13. The Htk ligand cDNA encodes a transmembrane protein of 336 amino acids. Binding competition experiments demonstrated a Kd of 535 pM for binding of Htk-Fc to the Htk ligand. Incubation of 3T3 cells expressing Htk with COS-7 cells expressing the ligand resulted in tyrosine phosphorylation of Htk. The ligand, like its receptor, is widely expressed and may function in a variety of tissues. However, we localized hematopoietic expression of Htk to the monocytic lineage, suggesting that the ligand may play a role in differentiation and/or proliferation of these cells.

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Selected References

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  1. Andres A. C., Reid H. H., Zürcher G., Blaschke R. J., Albrecht D., Ziemiecki A. Expression of two novel eph-related receptor protein tyrosine kinases in mammary gland development and carcinogenesis. Oncogene. 1994 May;9(5):1461–1467. [PubMed] [Google Scholar]
  2. Bartley T. D., Hunt R. W., Welcher A. A., Boyle W. J., Parker V. P., Lindberg R. A., Lu H. S., Colombero A. M., Elliott R. L., Guthrie B. A. B61 is a ligand for the ECK receptor protein-tyrosine kinase. Nature. 1994 Apr 7;368(6471):558–560. doi: 10.1038/368558a0. [DOI] [PubMed] [Google Scholar]
  3. Beckmann M. P., Cerretti D. P., Baum P., Vanden Bos T., James L., Farrah T., Kozlosky C., Hollingsworth T., Shilling H., Maraskovsky E. Molecular characterization of a family of ligands for eph-related tyrosine kinase receptors. EMBO J. 1994 Aug 15;13(16):3757–3762. doi: 10.1002/j.1460-2075.1994.tb06685.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bennett B. D., Wang Z., Kuang W. J., Wang A., Groopman J. E., Goeddel D. V., Scadden D. T. Cloning and characterization of HTK, a novel transmembrane tyrosine kinase of the EPH subfamily. J Biol Chem. 1994 May 13;269(19):14211–14218. [PubMed] [Google Scholar]
  5. Cheng H. J., Flanagan J. G. Identification and cloning of ELF-1, a developmentally expressed ligand for the Mek4 and Sek receptor tyrosine kinases. Cell. 1994 Oct 7;79(1):157–168. doi: 10.1016/0092-8674(94)90408-1. [DOI] [PubMed] [Google Scholar]
  6. Davis S., Gale N. W., Aldrich T. H., Maisonpierre P. C., Lhotak V., Pawson T., Goldfarb M., Yancopoulos G. D. Ligands for EPH-related receptor tyrosine kinases that require membrane attachment or clustering for activity. Science. 1994 Nov 4;266(5186):816–819. doi: 10.1126/science.7973638. [DOI] [PubMed] [Google Scholar]
  7. Gearing D. P., King J. A., Gough N. M., Nicola N. A. Expression cloning of a receptor for human granulocyte-macrophage colony-stimulating factor. EMBO J. 1989 Dec 1;8(12):3667–3676. doi: 10.1002/j.1460-2075.1989.tb08541.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Holmes W. E., Lee J., Kuang W. J., Rice G. C., Wood W. I. Structure and functional expression of a human interleukin-8 receptor. Science. 1991 Sep 13;253(5025):1278–1280. doi: 10.1126/science.1840701. [DOI] [PubMed] [Google Scholar]
  9. Jordan C. T., McKearn J. P., Lemischka I. R. Cellular and developmental properties of fetal hematopoietic stem cells. Cell. 1990 Jun 15;61(6):953–963. doi: 10.1016/0092-8674(90)90061-i. [DOI] [PubMed] [Google Scholar]
  10. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  11. Lee J., Horuk R., Rice G. C., Bennett G. L., Camerato T., Wood W. I. Characterization of two high affinity human interleukin-8 receptors. J Biol Chem. 1992 Aug 15;267(23):16283–16287. [PubMed] [Google Scholar]
  12. Lhoták V., Greer P., Letwin K., Pawson T. Characterization of elk, a brain-specific receptor tyrosine kinase. Mol Cell Biol. 1991 May;11(5):2496–2502. doi: 10.1128/mcb.11.5.2496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lhoták V., Pawson T. Biological and biochemical activities of a chimeric epidermal growth factor-Elk receptor tyrosine kinase. Mol Cell Biol. 1993 Nov;13(11):7071–7079. doi: 10.1128/mcb.13.11.7071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lieschke G. J., Stanley E., Grail D., Hodgson G., Sinickas V., Gall J. A., Sinclair R. A., Dunn A. R. Mice lacking both macrophage- and granulocyte-macrophage colony-stimulating factor have macrophages and coexistent osteopetrosis and severe lung disease. Blood. 1994 Jul 1;84(1):27–35. [PubMed] [Google Scholar]
  15. MacKay K., Striker L. J., Elliot S., Pinkert C. A., Brinster R. L., Striker G. E. Glomerular epithelial, mesangial, and endothelial cell lines from transgenic mice. Kidney Int. 1988 Mar;33(3):677–684. doi: 10.1038/ki.1988.53. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
  18. Pawson T., Bernstein A. Receptor tyrosine kinases: genetic evidence for their role in Drosophila and mouse development. Trends Genet. 1990 Nov;6(11):350–356. doi: 10.1016/0168-9525(90)90276-c. [DOI] [PubMed] [Google Scholar]
  19. Sajjadi F. G., Pasquale E. B. Five novel avian Eph-related tyrosine kinases are differentially expressed. Oncogene. 1993 Jul;8(7):1807–1813. [PubMed] [Google Scholar]
  20. Schlessinger J., Ullrich A. Growth factor signaling by receptor tyrosine kinases. Neuron. 1992 Sep;9(3):383–391. doi: 10.1016/0896-6273(92)90177-f. [DOI] [PubMed] [Google Scholar]
  21. Shao H., Lou L., Pandey A., Pasquale E. B., Dixit V. M. cDNA cloning and characterization of a ligand for the Cek5 receptor protein-tyrosine kinase. J Biol Chem. 1994 Oct 28;269(43):26606–26609. [PubMed] [Google Scholar]
  22. Smith C. A., Gruss H. J., Davis T., Anderson D., Farrah T., Baker E., Sutherland G. R., Brannan C. I., Copeland N. G., Jenkins N. A. CD30 antigen, a marker for Hodgkin's lymphoma, is a receptor whose ligand defines an emerging family of cytokines with homology to TNF. Cell. 1993 Jul 2;73(7):1349–1360. doi: 10.1016/0092-8674(93)90361-s. [DOI] [PubMed] [Google Scholar]
  23. Talhouk R. S., Bissell M. J., Werb Z. Coordinated expression of extracellular matrix-degrading proteinases and their inhibitors regulates mammary epithelial function during involution. J Cell Biol. 1992 Sep;118(5):1271–1282. doi: 10.1083/jcb.118.5.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Urdal D. L., Call S. M., Jackson J. L., Dower S. K. Affinity purification and chemical analysis of the interleukin-1 receptor. J Biol Chem. 1988 Feb 25;263(6):2870–2877. [PubMed] [Google Scholar]
  25. Zeigler F. C., Bennett B. D., Jordan C. T., Spencer S. D., Baumhueter S., Carroll K. J., Hooley J., Bauer K., Matthews W. Cellular and molecular characterization of the role of the flk-2/flt-3 receptor tyrosine kinase in hematopoietic stem cells. Blood. 1994 Oct 15;84(8):2422–2430. [PubMed] [Google Scholar]
  26. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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