Identification of novel p53-binding proteins by biomolecular interaction analysis combined with tandem mass spectrometry.

Electrospray tandem mass spectrometry (ESI-MS/MS) was combined with biomolecular interaction analysis (BIA) to develop a method of direct protein identification after real-time analysis of protein protein interactions. Using this method, called BIA-MS/MS, we detected multiple p53-interacting proteins in whole tissue extracts from human placenta and liver. Peptide sequencing revealed three ...
proteins whose interaction with p53 had not been previously reported: a cyclin-dependent kinase inhibitor p57/Kip2, a serine/threonine protein phosphatase PP1C, and hemoglobin. Using our system, unambiguous sequence information can be obtained at the femto- to picomole level after repeating the recovery procedure five times. Furthermore, the association and dissociation constants are easily determined by kinetic analysis. This system provides a powerful tool for analyzing complex biological materials in a simple but highly specific and sensitive manner.
Mesh Terms:
Amino Acid Sequence, Brain, Brain Chemistry, Feasibility Studies, Female, Hemoglobins, Humans, Kinetics, Liver, Microchemistry, Molecular Sequence Data, Nanotechnology, Placenta, Pregnancy, Protein Binding, Sequence Analysis, Protein, Spectrometry, Mass, Electrospray Ionization, Surface Plasmon Resonance, Tumor Suppressor Protein p53
Mol. Biotechnol.
Date: Mar. 01, 2003
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