Use of electrospray ionization mass spectrometry and tandem mass spectrometry to study binding of F-0 inhibitors to ceroid lipofuscinosis protein, a model system for subunit c of mitochondrial ATP synthase
UNSPECIFIED. (1996) Use of electrospray ionization mass spectrometry and tandem mass spectrometry to study binding of F-0 inhibitors to ceroid lipofuscinosis protein, a model system for subunit c of mitochondrial ATP synthase. RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 10 (7). pp. 790-796. ISSN 0951-4198Full text not available from this repository.
Ceroid lipofuscinosis protein (CLP), the major accumulating protein in several forms of ceroid lipofuscinosis, has an amino acid sequence that is identical to that of the F-0 subunit c of normal bovine ATP synthase, Electrospray ionization mass spectrometry (ESI-MS) has shown that ovine CLP and normal bovine F-0 subunit c are identical, including a 42 mass unit post-translational modification, Although the identity and the location of this modification have not been fully established in both species, CLP can be used as a convenient and a unique source of subunit c for studies of F-0 inhibitor interactions by ESI-MS analysis.
Analysis of mixtures of CLP incubated with several known F-0 inhibitors showed that N,N'-dicyclohexyl-carbodiimide and organotins bind covalently to CLP but interactions with oligomycin and venturicidin were not observed, The sulphydryl inhibitors, 2,3-dimethoxy-5-methyl-1,4,-benzoquinone (UQ(0)) and N-ethyl maleimide (NEM) were also shown to bind covalently to the protein, The binding stoichiometry and the relative rate of reaction were then determined for each inhibitor. Tandem mass spectrometry experiments performed on the [M+5H](5+) ion of the intact CLP and of the complexes UQ(0)-CLP and NEM-CLP snowed the identification of 80% of the CLP sequence and revealed that UQ(0) and NEM are both bound to cysteine-64, This work shows the exceptional utility of ESI-MS in studies of the interaction of CLP with a range of inhibitors which are applicable to studies of the F-0 component of ATP synthase.
|Item Type:||Journal Article|
|Subjects:||Q Science > QD Chemistry
Q Science > QC Physics
|Journal or Publication Title:||RAPID COMMUNICATIONS IN MASS SPECTROMETRY|
|Publisher:||JOHN WILEY & SONS LTD|
|Number of Pages:||7|
|Page Range:||pp. 790-796|
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