Pharmacology; Phosphorylation; Protein Kinases; Signal Transduction; Crystallography, X-Ray; Receptor Protein-Tyrosine Kinases; Cell Proliferation; Drug Discovery
Center for X-Linked Hypophosphatemia, Yale
Our laboratory is exploring the mode of action of growth factor receptors and the intracellular signaling pathways that are activated in response to growth factor stimulation. Receptor tyrosine kinases and cytoplasmic protein tyrosine kinases play a critical role in the control of many cellular processes including: cell proliferation, differentiation, metabolism, as well as cell survival and cell migration. Various diseases are caused by dysfunctions in receptor tyrosine kinases or in critical components of signaling pathways that are activated by receptors tyrosine kinases. In addition, various developmental disorders are caused by loss of function mutations in receptor tyrosine kinases. We are using biochemical and genetic approaches as well as X-ray crystallography to determine the mechanism of activation of receptor tyrosine kinases, how they recruit target proteins, and the mode of action of downstream signaling proteins in normal cellular processes and in diseases caused by dysfunctional receptor tyrosine kinases.
Specialized Terms: Growth factor receptors; Intracellular signaling pathways; Protein kinases; Phosphorylation; SH2, SH3 and other protein modules involved in signal transduction; Drug discovery
Extensive Research Description
Tyrosine phosphorylation plays a critical role in the control of many cellular processes including cell proliferation, differentiation, metabolism, as well as cell survival and migration. Receptor tyrosine kinases undergo ligand dependent dimerization which activates their intrinsic protein tyrosine kinase (PTK) domains. We have determined the crystal structure of Stem cell factor (SCF) and fibroblast growth factor (FGF), two ligands of receptor tyrosine kinases. In addition, we have determined the crystal structure of FGF in complex with the extracellular ligand binding domain of FGF-receptor (FGFR) and with a heparin sulfate oligosacchride. The structure of the ternary FGF/heparin/FGFR complex provides a molecular view of how FGF acts in concert with heparin to induce the dimerization and activation of FGF-receptors. We have also determined the crystal structure of the catalytic PTK domain of FGFR in complex with an ATP analogue or in complex with specific PTK inhibitors of FGFR activity and function. These structures enabled the development of new specific inhibitor for PTKs that are currently being tested in clinical trials. Receptor tyrosine kinases undergo ligand-dependent dimerization, which activates their intrinsic protein tyrosine kinase activity resulting in autophosphorylation and subsequent interaction and recruitment of multiple cellular target proteins. The phosphorylated tyrosine residues together with their immediate flanking sequences function as binding sites for signaling molecules containing src homology 2 (SH2) domains. Many signaling proteins carry SH2 domains plus one or more small protein modules such as SH3, PH, PTB, WW or FYVE domains. These protein modules function as mediator of protein-protein or protein-lipid interactions that are critical for signal transmission. In addition to direct recruitment by RTKs, many signaling proteins are recruited by an alternative mechanism involving a family of membrane linked docking proteins such as FRS-2a, and b, IRS-1 and 2, and Gab-1 and 2, among many others. Recruitment of signaling proteins by RTKs or by docking proteins leads to activation of multiple signaling pathways resulting in stimulation of a variety of cellular responses. The small adapter protein Grb2, for example, is bound through its SH3 domains to short, proline-rich sequences in the carboxy terminal tail of the guanine nucleotide-releasing factor Sos. Interaction between Grb2 and Sos with tyrosine phosphorylated RTKs or docking proteins results in translocation of Sos to the plasma membrane allowing the exchange of GDP for GTP on Ras. The activated GTP-bound form of Ras then starts a kinase cascade composed of Raf, MAPKK, and MAPK leading to phosphorylation of prooncogene Jun on serine and threonine residues to induce transcriptional activation. These and other signaling pathways that are activated by RTKs regulate multiple cellular processes. Many cancers and other diseases are caused by dysfunctions in RTKs or in components of their intracellular pathways.
- Structural analysis of the mechanism of phosphorylation of a critical autoregulatory tyrosine residue in FGFR1 kinase domain. Kobashigawa Y, Amano S, Yokogawa M, Kumeta H, Morioka H, Inouye M, Schlessinger J, Inagaki F. Genes Cells. 2015 Aug 24. doi: 10.1111/gtc.12277. [Epub ahead of print]
- Exome sequencing identifies recurrent mutations in NF1 and RASopathy genes in sun-exposed melanomas. Krauthammer M, Kong Y, Bacchiocchi A, Evans P, Pornputtapong N, Wu C, McCusker JP, Ma S, Cheng E, Straub R, Serin M, Bosenberg M, Ariyan S, Narayan D, Sznol M, Kluger HM, Mane S, Schlessinger J, Lifton RP, Halaban R. Nat Genet. 2015 Sep;47(9):996-1002. doi: 10.1038/ng.3361. Epub 2015 Jul 27.
- FGF1 and FGF19 reverse diabetes by suppression of the hypothalamic-pituitary-adrenal axis. Perry RJ, Lee S, Ma L, Zhang D, Schlessinger J, Shulman GI. Nat Commun. 2015 Apr 28;6:6980. doi: 10.1038/ncomms7980.
- Heparin is an activating ligand of the orphan receptor tyrosine kinase ALK. Murray PB, Lax I, Reshetnyak A, Ligon GF, Lillquist JS, Natoli EJ Jr, Shi X, Folta-Stogniew E, Gunel M, Alvarado D, Schlessinger J. Sci Signal. 2015 Jan 20;8(360):ra6. doi: 10.1126/scisignal.2005916.
- Whole-exome sequencing characterizes the landscape of somatic mutations and copy number alterations in adrenocortical carcinoma. Juhlin CC, Goh G, Healy JM, Fonseca AL, Scholl UI, Stenman A, Kunstman JW, Brown TC, Overton JD, Mane SM, Nelson-Williams C, Bäckdahl M, Suttorp AC, Haase M, Choi M, Schlessinger J, Rimm DL, Höög A, Prasad ML, Korah R, Larsson C, Lifton RP, Carling T. J Clin Endocrinol Metab. 2015 Mar;100(3):E493-502. doi: 10.1210/jc.2014-3282. Epub 2014 Dec 9.
- Differential TAM receptor-ligand-phospholipid interactions delimit differential TAM bioactivities. Lew ED, Oh J, Burrola PG, Lax I, Zagórska A, Través PG, Schlessinger J, Lemke G. Elife. 2014 Sep 29;3. doi: 10.7554/eLife.03385.
- The docking protein FRS2α is a critical regulator of VEGF receptors signaling. Chen PY, Qin L, Zhuang ZW, Tellides G, Lax I, Schlessinger J, Simons M. Proc Natl Acad Sci U S A. 2014 Apr 15;111(15):5514-9. doi: 10.1073/pnas.1404545111. Epub 2014 Apr 2.
- Receptor tyrosine kinases: legacy of the first two decades. Schlessinger J. Cold Spring Harb Perspect Biol. 2014 Mar 1;6(3). pii: a008912. doi: 10.1101/cshperspect.a008912. Review.
- Structure, domain organization, and different conformational states of stem cell factor-induced intact KIT dimers. Opatowsky Y, Lax I, Tomé F, Bleichert F, Unger VM, Schlessinger J. Proc Natl Acad Sci U S A. 2014 Feb 4;111(5):1772-7. doi: 10.1073/pnas.1323254111. Epub 2014 Jan 21.
- Structural basis for KIT receptor tyrosine kinase inhibition by antibodies targeting the D4 membrane-proximal region. Reshetnyak AV, Nelson B, Shi X, Boggon TJ, Pavlenco A, Mandel-Bausch EM, Tome F, Suzuki Y, Sidhu SS, Lax I, Schlessinger J. Proc Natl Acad Sci U S A. 2013 Oct 29;110(44):17832-7. doi: 10.1073/pnas.1317118110. Epub 2013 Oct 14.
- Landscape of somatic single-nucleotide and copy-number mutations in uterine serous carcinoma. Zhao S, Choi M, Overton JD, Bellone S, Roque DM, Cocco E, Guzzo F, English DP, Varughese J, Gasparrini S, Bortolomai I, Buza N, Hui P, Abu-Khalaf M, Ravaggi A, Bignotti E, Bandiera E, Romani C, Todeschini P, Tassi R, Zanotti L, Carrara L, Pecorelli S, Silasi DA, Ratner E, Azodi M, Schwartz PE, Rutherford TJ, Stiegler AL, Mane S, Boggon TJ, Schlessinger J, Lifton RP, Santin AD. Proc Natl Acad Sci U S A. 2013 Feb 19;110(8):2916-21. doi: 10.1073/pnas.1222577110. Epub 2013 Jan 28.
- RAC1P29S is a spontaneously activating cancer-associated GTPase. Davis MJ, Ha BH, Holman EC, Halaban R, Schlessinger J, Boggon TJ. Proc Natl Acad Sci U S A. 2013 Jan 15;110(3):912-7. doi: 10.1073/pnas.1220895110. Epub 2013 Jan 2.
- M.A. Lemmon and J. Schlessinger. Cell signaling by receptor tyrosine kinases. Cell 141, 1117-1134 (2010).
- J.H. Bae, T.J. Boggon, F. Tomé, V. Mandiyan, I. Lax, and J. Schlessinger. An asymmetric receptor contact is required for tyrosine autophosphorylation of fibroblast growth factor receptor in living cells. Proc. Natl. Acad. Sci. USA 107, 2866-2871 (2010).
- J.H. Bae, E.D. Lew, S. Yuzawa, F. Tome, I. Lax, and J. Schlessinger. The selectivity of receptor tyrosine kinase signaling is controlled by a secondary SH2 domain binding site. Cell 138, 514-524 (2009).
- E.D. Lew, Jae-Hyun Bae, E. Rohmann, B. Wollnik and J. Schlessinger. Structural basis for reduced FGFR2 activity in LADD Syndrome: Implications for FGFR autoinhibition and activation. Proc. Natl. Acad. Sci. USA 140, 19802-19807 (2007).
- S. Yuzawa, Y. Opatowsky, Z. Zhang, V. Mandiyan, I. Lax, and J. Schlessinger. Structural basis for activation of the receptor tyrosine kinase Kit by stem cell factor. Cell 130, 323-334 (2007).