PMID:15459336
| Citation |
Gupta, R, Hamdan, SM, Dixon, NE, Sheil, MM and Beck, JL (2004) Application of electrospray ionization mass spectrometry to study the hydrophobic interaction between the epsilon and theta subunits of DNA polymerase III. Protein Sci. 13:2878-87 |
|---|---|
| Abstract |
The interactions between the N-terminal domain of the epsilon (epsilon186) and theta subunits of DNA polymerase III of Escherichia coli were investigated using electrospray ionization mass spectrometry. The epsilon186-theta complex was stable in 9 M ammonium actetate (pH 8), suggesting that hydrophobic interactions have a predominant contribution to the stability of the complex. Addition of primary alkanols to epsilon186-theta in 0.1 M ammonium acetate (pH 8), led to dissociation of the complex, as observed in the mass spectrometer. The concentrations of methanol, ethanol, and 1-propanol required to dissociate 50% of the complex were 8.9 M, 4.8 M, and 1.7 M, respectively. Closer scrutiny of the effect of alkanols on epsilon186, theta, and epsilon186-theta showed that epsilon186 formed soluble aggregates prior to precipitation, and that the association of epsilon186 with theta stabilized epsilon186. In-source collision-induced dissociation experiments and other results suggested that the epsilon186-theta complex dissociated in the mass spectrometer, and that the stability (with respect to dissociation) of the complex in vacuo was dependent on the solution from which it was sampled. |
| Links |
PubMed PMC2286593 Online version:10.1110/ps.04889604 |
| Keywords |
DNA Polymerase III/chemistry; Enzyme Stability; Escherichia coli Proteins/chemistry; Hydrophobic and Hydrophilic Interactions; Protein Structure, Quaternary; Protein Subunits/chemistry; Spectrometry, Mass, Electrospray Ionization/methods; Static Electricity |
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