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A novel subfractionation approach for mitochondrial proteins: A three‐dimensional mitochondrial proteome map
Author(s) -
Hanson Bonnie J.,
Schulenberg Birte,
Patton Wayne F.,
Capaldi Roderick A.
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683()22:5<950::aid-elps950>3.0.co;2-d
Subject(s) - proteome , isoelectric focusing , mitochondrion , organelle , biology , gel electrophoresis , proteomics , two dimensional gel electrophoresis , biochemistry , computational biology , microbiology and biotechnology , enzyme , gene
As mitochondria play critical roles in both cell life and cell death, there is great interest in obtaining a human mitochondrial proteome map. Such a map could potentially be useful in diagnosing diseases, identifying targets for drug therapy, and in screening for unwanted drug side effects. In this paper, we present a novel approach to obtaining a human mitochondrial proteome map that combines sucrose gradient centrifugation with standard two‐dimensional gel electrophoresis. The resulting three‐dimensional separation of proteins allows us to address some of the problems encountered during previous attempts to obtain mitochondrial proteome maps such as resolution of proteins and solubility of hydrophobic proteins during isoelectric focusing. In addition, we show that this new approach provides functional information about protein complexes within the organelle that is not obtained with two‐dimensional gel electrophoresis of whole mitochondria.