The use of 2-D gels to identify novel protein-protein interactions in the cochlea.
Functional proteomics comprises a wide range of technologies for the identification of novel protein-protein interactions and biological markers. Studies of protein-protein interactions have gained from the development of techniques and technologies such as immunoprecipitation, preparative two-dimensional (2-D) gel electrophoresis for peptide mass fingerprinting (PMF), using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry, and liquid chromatography-tandem mass spectrometry (LC-MS/MS). These applications enabled the discovery of putative protein partners without a priori knowledge of which one(s) might be relevant. Here, we report the methods by which membrane proteins are isolated from cochlear tissues and prepared for identification by mass spectrometry techniques.
Department of Otolaryngology-HNS, University of South Florida College of Medicine, Tampa, FL, USA.
Thandavarayan
T
Kathiresan
Margaret C
MC
Harvey
Bernd H A
BH
Sokolowski
D000818:Animals; D003051:Cochlea / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D047468:Immunoprecipitation; D007525:Isoelectric Focusing; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism
D014357:Trypsin