Analysis of Individual Signaling Complexes by mMAPS, a Flow-Proteometric System

Curr Protoc Mol Biol. 2016 Apr 1:114:20.11.1-20.11.22. doi: 10.1002/0471142727.mb2011s114.

Abstract

Signal transduction is essential for maintaining normal cell physiological functions, and deregulation of signaling can lead to diseases such as diabetes and cancers. Some of the major players in signal delivery are molecular complexes composed of proteins and nucleic acids. This unit describes a technique called microchannel for multiparameter analysis of proteins in a single complex (mMAPS) for analyzing and quantifying individual target signaling complexes. mMAPS is a flow-proteometric system that allows detection of individual proteins or complexes flowing through a microfluidic channel. Specific target proteins and nucleic acids labeled by fluorescent tags are harvested from tissues or cultured cells for analysis by the mMAPS system. Overall, mMAPS enables both detection of multiple components within a single complex and direct quantification of different populations of molecular complexes in one setting in a short timeframe and requiring very low sample input.

Keywords: flow proteometry; microfluidic; protein-protein interaction; protein−nucleic acid interaction; single complex; single molecule.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calibration
  • Cells, Cultured
  • Humans
  • Microfluidic Analytical Techniques*
  • Proteins* / chemistry
  • Proteins* / metabolism
  • Proteomics / instrumentation
  • Proteomics / methods*
  • Quality Control

Substances

  • Proteins