PMID:15850376
| Citation |
Natale, P, den Blaauwen, T, van der Does, C and Driessen, AJ (2005) Conformational state of the SecYEG-bound SecA probed by single tryptophan fluorescence spectroscopy. Biochemistry 44:6424-32 |
|---|---|
| Abstract |
The SecYEG complex is a membrane-embedded channel that permits the passage of precursor proteins (preproteins) across the inner membrane of Escherichia coli. SecA is a molecular motor that associates with the SecYEG pore and drives the stepwise translocation of preproteins across the membrane through multiple cycles of ATP binding and hydrolysis. We have investigated the conformational state of soluble and SecYEG-bound SecA using single tryptophan mutants of SecA. The fluorescence spectral properties of the single tryptophans of SecA and their accessibility to the quencher acrylamide demonstrate that SecA undergoes a conformational change that results in a more compact structure upon binding of ATP and binding to the SecYEG pore. In addition, SecYEG-bound SecA undergoes ATP-dependent conformational changes that are not observed for soluble SecA. These data support a model in which binding to the SecYEG channel has a major impact on the SecA conformation. |
| Links |
PubMed Online version:10.1021/bi047488r |
| Keywords |
Acrylamide/chemistry; Adenosine Triphosphatases/chemistry; Adenosine Triphosphatases/genetics; Adenosine Triphosphatases/metabolism; Amino Acid Substitution/genetics; Bacterial Proteins/chemistry; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Genetic Complementation Test; Membrane Proteins/chemistry; Membrane Proteins/genetics; Membrane Proteins/metabolism; Membrane Transport Proteins/chemistry; Membrane Transport Proteins/genetics; Membrane Transport Proteins/metabolism; Protein Binding/genetics; Protein Conformation; Protein Transport/genetics; Solubility; Spectrometry, Fluorescence/methods; Temperature; Tryptophan/chemistry; Tryptophan/genetics |
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