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Takanari Inoue, Ph.D.

Takanari Inoue, Ph.D.

Academic Titles: 
Associate Professor
Position Title: 
Principal Investigator
Appointments/Affiliations: 
443-287-7668 (office)
443-287-7669 (lab)

Department of Cell Biology
Johns Hopkins University School of Medicine
855 N. Wolfe Street, 453 Rangos
Baltimore, MD 21205

Research Interest: 
Directed cell migration; tumor metastasis; primary cilia; synthetic chemical biology; technology development

Research Topic: Synthetic Cell Biology

Our research focuses on synthetic cell biology to dissect intricate signaling networks. In particular, we investigate positive-feedback mechanisms underlying the initiation of neutrophil chemotaxis (known as a symmetry breaking process), as well as spatio-temporally compartmentalized Ras signaling. In parallel, our lab also tries to understand how cell morphology affects biochemical functions in cells. Ultimately, we will generate completely orthogonal nano-machinery in cells that can achieve existing, and even novel, cellular functions. [Inoue1.jpg] 

 

 

 

Figure 1: Engineering cellular morphology. Time series of confocal fluorescence images of NIH3T3 fibroblasts experiencing striking morphological changes upon rapid activation of small GTPases such as Rac, Cdc42, and RhoA.

 

 

 

 

Figure 2: Inducible cell migration. Time series of confocal dual-color fluorescence images of differentiated HL-60 cells in which PI3K or Rac is rapidly and inducibly activated without activating any molecules upstream. Such synthetic activation of PI3K, but not Rac, led to cell polarization and following migration. Yellow: PI3K or Rac activated cells, Blue: Nuclei.

 

 


Figure 3: Tools to manipulate protein activity at specific subcellular compartments (a) A principle diagram for inducible recruitment of signaling molecules (green circles). c, cytoplasm, o, organelle. (b) Confocal fluorescence images of HeLa cells showing rapid translocation of cytoplasmic proteins to specific organelles upon 5 M iRap addition for 5 minutes. Scale bar indicates 20 m.

Biosketch: 
Lab Members:
Namesort descending Classification Email Phone
Allison Suarez Student asuarez4@jhmi.edu 443-287-7669
Robert DeRose Research Specialist jderose1@jhmi.edu 443-287-7669
Shiva Razavi Student srazavi@jhmi.edu 443-287-7669
Siew Cheng Phua Student sphua1@jhmi.edu 443-287-7669
Steven Su Student ssu7@jhu.edu 443-287-7669
Yu-Chun (Frank) Lin, Ph.D. Postdoctoral Fellow ylin54@jhmi.edu 443-287-7669
Yuchuan Miao Student ymiao1@jhmi.edu 410-955-4699
Selected Publications:
Onuma H, Komatsu T, Arita M, Hanaoka K, Ueno T, Terai T, Nagano T, Inoue T.  2014.  Rapidly rendering cells phagocytic through a cell surface display technique and concurrent Rac activation.  Sci Signal.  Jul 15;7(334):rs4. doi: 10.1126/scisignal.2005123.
Suarez A., Ueno T., Huebner R., McCaffery J.M., and Inoue T.* “Bin/Amphiphysin/Rvs (BAR) family members bend membranes in cells” Scientific Reports 4, 4693 (2014)  
Kobayashi T., Kim S., Lin Y.C., Inoue T., and Dynlacht B.D. “CP110-interacting proteins, Talpid3 and Cep290, play overlapping and distinct roles in cilia assembly.” Journal of Cell Biology 204, 215 (2014) 
Su S., Phua S.C., DeRose R., Chiba S., Narita K., Kalugin P.N., Katada T., Kontani K., Takeda S. and Inoue T. “Genetically encoded calcium indicator illuminates calcium dynamics in cilia.” Nature Methods 10, 1105 (2013)
Thevathasan, J.V., Tan E., Hui Z., Lin Y.C., Li Y., Inoue T. and Fivaz M. “The small GTPase HRas shapes local PI3K signals through positive feedback and regulates persistent membrane extension in migrating fibroblasts.” Molecular Biology of the Cell 24, 2228 (2013)  
Lin Y.C., Niewiadomski P., Lin B., Nakamura H., Phua S.C., Jiao J., Levchenko A., Inoue T., Rohatgi T., and Inoue T. “Chemically-inducible diffusion trap reveals molecular sieve-like barrier at primary cilia“ Nature Chemical Biology 9, 437 (2013)
Lin YC, Liu T-Y, Razavi S. and Inoue T. “Rapidly Reversible Manipulation of Molecular Activities Using Dual Chemical Dimerizers” Angewandte Chemie 52, 6450 (2013)
Thevathasan, J.V., Tan E., Hui Z., Lin Y.C., Li Y., Inoue T. and Fivaz M. “Local positive feedback from PI3K to Ras drives cell polarization and migration” Molecular Biology of the Cell (doi:10.1091/mbc.E12-12-0905)
Miyamoto T., DeRose R., Suarez A., Ueno T., Chen M., Sun T.-p., Wolfgang M.J., Mukherjee C., Meyers D. and Inoue T. “Generation of Intracellular Logic Gates with Two Orthogonal Chemically Inducible Systems” Nature Chemical Biology (Accepted 2012)
Ueno T., Falkenburger B.H., Pohlmeyer C., and Inoue T. “Triggering Actin Comets Versus Membrane Ruffles: Distinctive Effects of Phosphoinositides on Actin Reorganization” ScienceSignaling 4(203), ra87 (2011) (Cover Article)
Umeda N., Ueno T., Pohlmeyer C., Nagano T. and Inoue T. “A photocleavable rapamycin conjugate for spatiotemporal control of small GTPase activity” Journal of American Chemical Society 133(1), 12-14 (2011) Note: “Scientist use light to move molecules within living cells” Science News article in ScienceDaily
Komatsu T., Kukelyansky I., McCaffery J.M., Ueno T., Varela L.C. and Inoue T. “Organelle-Specific, Rapid Induction of Molecular Activities and Membrane Tethering” Nature Methods 7, 206-208 (2010) Note: “Hopkins researchers put proteins right where they want them” Breaking News article in Genetic Engineering and Biotechnology News
Rahdar M., Inoue T., Meyer T., Zhang J., Vazquez F., and Devreotes P.N. “A phosphorylation-dependent intramolecular interaction regulates the membrane association and activity of the tumor suppressor PTEN” Proc. Natl. Acad. Sci. U S A. 106(2):480-5 (2009)
Inoue T. and Meyer T. “Synthetic activation of endogenous PI3K and Rac identifies an AND-gate switch for cell polarization and migration” PLoS ONE 3(8), e3068 (2008)
Fivaz M., Bandara S., Inoue T. and Meyer T. "Robust neuronal symmetry breaking by Ras-triggered local positive feedback" Current Biology 18, 44-50 (2008)
Suh BC*, Inoue T*, Meyer T, and Hille B. “Rapid chemically-induced changes of PtdIns(4,5)P2 gate KCNQ ion channels” Science 314, 1454-1457 (2006) (*Contributed Equally) Note: “Perspectives” (Science 314, 1402-1403 (2006)), “Editor’s Choice” (Science STKE 364, tw410 (2006)), “Spotlight” (ACS Chem. Biol. 1, 608 (2006)), “Research Highlights” (Nature Methods 4, 7 (2007))
Heo WD, Inoue T, Park WS, Kim ML, Park BO, Wandless TJ, and Meyer T. “PI(3,4,5)P3 and PI(4,5)P2 lipids target Ras, Rho, Arf and Rab GTPases to the plasma membrane” Science 314, 1458-1461 (2006)
Inoue T, Heo WD, Grimley JS, Wandless TJ, and Meyer T. “Inducible translocation strategies to rapidly activate and inhibit small GTPase signaling pathways” Nature Methods 2, 415-418 (2005)
Inoue T, Kikuchi K, Hirose K, Iino M, and Nagano T. “Spatiotemporal Laser Inactivation of Inositol 1,4,5-Trisphosphate Receptors Using Synthetic Small-molecule Probes” Chem. Biol. 10, 503-509 (2003) Note: “Cover art”
Inoue T, Kikuchi K, Hirose K, Iino M, and Nagano T. “Small molecule-based laser inactivation of inositol 1,4,5-trisphosphate receptor” Chem. Biol. 8, 9-15 (2001)