PMID:15919996
| Citation |
Daley, DO, Rapp, M, Granseth, E, Melén, K, Drew, D and von Heijne, G (2005) Global topology analysis of the Escherichia coli inner membrane proteome. Science 308:1321-3 |
|---|---|
| Abstract |
The protein complement of cellular membranes is notoriously resistant to standard proteomic analysis and structural studies. As a result, membrane proteomes remain ill-defined. Here, we report a global topology analysis of the Escherichia coli inner membrane proteome. Using C-terminal tagging with the alkaline phosphatase and green fluorescent protein, we established the periplasmic or cytoplasmic locations of the C termini for 601 inner membrane proteins. By constraining a topology prediction algorithm with this data, we derived high-quality topology models for the 601 proteins, providing a firm foundation for future functional studies of this and other membrane proteomes. We also estimated the overexpression potential for 397 green fluorescent protein fusions; the results suggest that a large fraction of all inner membrane proteins can be produced in sufficient quantities for biochemical and structural work. |
| Links |
PubMed Online version:10.1126/science.1109730 |
| Keywords |
Alkaline Phosphatase/analysis; Alkaline Phosphatase/genetics; Cell Membrane/chemistry; Cloning, Molecular; Computational Biology; Cytoplasm/chemistry; Escherichia coli/chemistry; Escherichia coli/genetics; Escherichia coli/ultrastructure; Escherichia coli Proteins/analysis; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/genetics; Escherichia coli Proteins/physiology; Gene Duplication; Genes, Bacterial; Green Fluorescent Proteins/analysis; Green Fluorescent Proteins/genetics; Membrane Proteins/analysis; Membrane Proteins/chemistry; Membrane Proteins/genetics; Membrane Proteins/physiology; Periplasm/chemistry; Protein Structure, Secondary; Proteome; Recombinant Fusion Proteins |
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