mMAPS: A Flow-Proteometric Technique to Analyze Protein-Protein Interactions in Individual Signaling Complexes

Sci. Signal., 4 March 2014
Vol. 7, Issue 315, p. rs1
DOI: 10.1126/scisignal.2004620

mMAPS: A Flow-Proteometric Technique to Analyze Protein-Protein Interactions in Individual Signaling Complexes

  1. Chao-Kai Chou1,2,
  2. Heng-Huan Lee1,3,*,
  3. Pei-Hsiang Tsou1,*,
  4. Chun-Te Chen1,2,
  5. Jung-Mao Hsu1,
  6. Hirohito Yamaguchi1,
  7. Ying-Nai Wang1,2,4,
  8. Hong-Jen Lee1,3,
  9. Jennifer L. Hsu1,2,
  10. Jin-Fong Lee1,
  11. Jun Kameoka5,, and
  12. Mien-Chie Hung1,2,3,4,
  1. 1Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.
  2. 2Graduate Institute for Cancer Biology and Center for Molecular Medicine, China Medical University, Taichung 404, Taiwan.
  3. 3Cancer Biology Program, Graduate School of Biomedical Sciences, The University of Texas Health Sciences Center at Houston, Houston, TX 77030, USA.
  4. 4Department of Biotechnology, Asia University, Taichung 413, Taiwan.
  5. 5Department of Electrical and Computer Engineering, Texas A&M University, College Station, TX 77843, USA.
  1. Corresponding author. E-mail: mhung{at}mdanderson.org (M.-C.H.); kameoka{at}mail.ece.tamu.edu (J.K.)
  • * These authors contributed equally to this work.

Abstract

Signal transduction is a dynamic process that regulates cellular functions through multiple types of biomolecular interactions, such as the interactions between proteins and between proteins and nucleic acids. However, the techniques currently available for identifying protein-protein or protein–nucleic acid complexes typically provide information about the overall population of signaling complexes in a sample instead of information about the individual signaling complexes therein. We developed a technique called “microchannel for multiparameter analysis of proteins in a single complex” (mMAPS) that simultaneously detected individual target proteins either singly or in a multicomponent complex in cell or tissue lysates. We detected the target proteins labeled with fluorophores by flow proteometry, which provided quantified data in the form of multidimensional fluorescence plots. Using mMAPS, we quantified individual complexes of epidermal growth factor (EGF) with its receptor EGFR, EGFR with signal transducer and activator of transcription 3 (STAT3), and STAT3 with the acetylase p300 and DNA in lysates from cultured cells with and without treatment with EGF, as well as in lysates from tumor xenograft tissue. Consistent with the ability of this method to reveal the dynamics of signaling protein interactions, we observed that cells treated with EGF induced the interaction of EGF with EGFR and the autophosphorylation of EGFR, but this interaction decreased with longer treatment time. Thus, we expect that this technique may reveal new aspects of molecular interaction dynamics.

Citation:

C.-K. Chou, H.-H. Lee, P.-H. Tsou, C.-T. Chen, J.-M. Hsu, H. Yamaguchi, Y.-N. Wang, H.-J. Lee, J. L. Hsu, J.-F. Lee, J. Kameoka, and M.-C. Hung, mMAPS: A Flow-Proteometric Technique to Analyze Protein-Protein Interactions in Individual Signaling Complexes. Sci. Signal. 7, rs1 (2014).
Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882