Activation and Regulation of the Understudied Kinome using MIB/MS Technology

• Gary L. Johnson, Ph.D.
  Kenan Distinguished Professor and Chair
  Department of Pharmacology

• Shawn Gomez, Ph.D.
  Associate Professor Department of Biomedical Engineering
  Adjunct Assistant Professor
  Department of Computer Science

Overview

Our work focuses on understanding the kinase protein class by identifying the activation or inhibition of kinases due to presence of perturbation and compare those states in known signaling networks. One rationale for these studies is the SKBR-3 kinome response to Lapatinib, which have shown to yield no difference, activation, or inhibition states of the kinome. And the critical component is the timing of response. Collaboration work with Pharmaceutical industry, including GSK and their ‘window trials’. Via these, we have gathered information on kinome reprogrammed and single disease have heterogeneity (among patient samples) with response to compound. We are working on developing a ‘kinome activation atlas’ by defining nodal kinases within understudied kinome.

Technology developed and utilized (showing in one of the pictures) is to capture native kinases via a column of Multiplexed Inhibitors Beads, containing kinase inhibitors that have broad binding characteristics. This enrichment of kinases are identified via Mass Spectrometry and the states of the kinase (activated or not) is assessed leading to a kinome activation signature.

Publications

Stuhlmiller, T.J., Miller, S.M., Zawistowski, Kazuhiro Nakamura, K., Beltran, A., Duncan, J.S., Collins, K.L.A., Granger, D.A., Rachel A. Reuther, R.A., Graves, L.M., Gomez, S.M., Kuan, P-F., Joel S. Parker, J.S., Chen, X., Sciaky, N., Carey, L.A., Shelton Earp, H.S., Jin, J., Johnson, G.L. (2015) Inhibition of Lapatinib-induced Kinome Reprogramming in ERBB2-positive Breast Cancer by Targeting BET Family Bromodomains, Cell Reports http://dx.doi.org/10.1016/j.celrep.2015.03.037

Links

• Gary Johnson’s lab: https://www.med.unc.edu/pharm/johnsonlab

• Shawn Gomez lab: http://gomezlab.bme.unc.edu/

NIH Funding Follow Link