title: Quantitative analysis of receptor tyrosine kinase-effector coupling at functionally relevant stimulus levels.
abstract: A major goal of current signaling research is to develop a quantitative understanding of how receptor activation is coupled to downstream signaling events and to functional cellular responses. Here, we measure how activation of the RET receptor tyrosine kinase on mouse neuroblastoma cells by the neurotrophin artemin (ART) is quantitatively coupled to key downstream effectors. We show that the efficiency of RET coupling to ERK and Akt depends strongly on ART concentration, and it is highest at the low (∼100 pM) ART levels required for neurite outgrowth. Quantitative discrimination between ERK and Akt pathway signaling similarly is highest at this low ART concentration. Stimulation of the cells with 100 pM ART activated RET at the rate of ∼10 molecules/cell/min, leading at 5-10 min to a transient peak of ∼150 phospho-ERK (pERK) molecules and ∼50 pAkt molecules per pRET, after which time the levels of these two signaling effectors fell by 25-50% while the pRET levels continued to slowly rise. Kinetic experiments showed that signaling effectors in different pathways respond to RET activation with different lag times, such that the balance of signal flux among the different pathways evolves over time. Our results illustrate that measurements using high, super-physiological growth factor levels can be misleading about quantitative features of receptor signaling. We propose a quantitative model describing how receptor-effector coupling efficiency links signal amplification to signal sensitization between receptor and effector, thereby providing insight into design principles underlying how receptors and their associated signaling machinery decode an extracellular signal to trigger a functional cellular outcome.
authors: From the Department of Chemistry, Boston University, Boston, Massachusetts 02215.
authors: Simin
authors: S
authors: Li
authors: From the Department of Chemistry, Boston University, Boston, Massachusetts 02215.
authors: Devayani
authors: D
authors: Bhave
authors: From the Department of Chemistry, Boston University, Boston, Massachusetts 02215.
authors: Jennifer M
authors: JM
authors: Chow
authors: From the Department of Chemistry, Boston University, Boston, Massachusetts 02215.
authors: Thomas V
authors: TV
authors: Riera
authors: From the Department of Chemistry, Boston University, Boston, Massachusetts 02215.
authors: Sandra
authors: S
authors: Schlee
authors: From the Department of Chemistry, Boston University, Boston, Massachusetts 02215.
authors: Simone
authors: S
authors: Rauch
authors: From the Department of Chemistry, Boston University, Boston, Massachusetts 02215.
authors: Mariya
authors: M
authors: Atanasova
authors: From the Department of Chemistry, Boston University, Boston, Massachusetts 02215.
authors: Richard L
authors: RL
authors: Cate
authors: From the Department of Chemistry, Boston University, Boston, Massachusetts 02215 whitty@bu.edu.
authors: Adrian
authors: A
authors: Whitty
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation; D007700:Kinetics; D008024:Ligands; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000235:genetics / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism* / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism*; D051096:Proto-Oncogene Proteins c-ret / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013997:Time Factors
chemical_list: C116599:Artn protein, mouse; D008024:Ligands; D009419:Nerve Tissue Proteins; D010750:Phosphoproteins; D051096:Proto-Oncogene Proteins c-ret; C494999:Ret protein, mouse; D051057:Proto-Oncogene Proteins c-akt; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3
keywords: Akt PKB; Artemin; Extracellular Signal-regulated Kinase (ERK); Kinetics; Mitogen-activated Protein Kinase (MAPK); Neurite Outgrowth; RET; Signal Gain; Signal Transduction; Systems Biology
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