ABCC1 p.Ser1137Cys
| Predicted by SNAP2: | A: D (59%), C: D (53%), D: D (85%), E: D (85%), F: D (75%), G: D (66%), H: D (80%), I: D (75%), K: D (85%), L: D (75%), M: D (66%), N: D (63%), P: D (91%), Q: D (80%), R: D (85%), T: D (63%), V: D (71%), W: D (91%), Y: D (80%), |
| Predicted by PROVEAN: | A: D, C: D, D: D, E: D, F: D, G: D, H: D, I: D, K: D, L: D, M: D, N: D, P: D, Q: D, R: D, T: D, V: D, W: D, Y: D, |
[switch to compact view]
Comments [show]
None has been submitted yet.
[hide] Dynamic ligand-induced conformational rearrangemen... J Biol Chem. 2012 Jan 6;287(2):1112-27. Epub 2011 Nov 15. Verhalen B, Ernst S, Borsch M, Wilkens S
Dynamic ligand-induced conformational rearrangements in P-glycoprotein as probed by fluorescence resonance energy transfer spectroscopy.
J Biol Chem. 2012 Jan 6;287(2):1112-27. Epub 2011 Nov 15., [PMID:22086917]
Abstract [show]
P-glycoprotein (Pgp), a member of the ATP-binding cassette transporter family, functions as an ATP hydrolysis-driven efflux pump to rid the cell of toxic organic compounds, including a variety of drugs used in anticancer chemotherapy. Here, we used fluorescence resonance energy transfer (FRET) spectroscopy to delineate the structural rearrangements the two nucleotide binding domains (NBDs) are undergoing during the catalytic cycle. Pairs of cysteines were introduced into equivalent regions in the N- and C-terminal NBDs for labeling with fluorescent dyes for ensemble and single-molecule FRET spectroscopy. In the ensemble FRET, a decrease of the donor to acceptor (D/A) ratio was observed upon addition of drug and ATP. Vanadate trapping further decreased the D/A ratio, indicating close association of the two NBDs. One of the cysteine mutants was further analyzed using confocal single-molecule FRET spectroscopy. Single Pgp molecules showed fast fluctuations of the FRET efficiencies, indicating movements of the NBDs on a time scale of 10-100 ms. Populations of low, medium, and high FRET efficiencies were observed during drug-stimulated MgATP hydrolysis, suggesting the presence of at least three major conformations of the NBDs during catalysis. Under conditions of vanadate trapping, most molecules displayed high FRET efficiency states, whereas with cyclosporin, more molecules showed low FRET efficiency. Different dwell times of the FRET states were found for the distinct biochemical conditions, with the fastest movements during active turnover. The FRET spectroscopy observations are discussed in context of a model of the catalytic mechanism of Pgp.
Comments [show]
None has been submitted yet.
No. Sentence Comment
65 Site-directed mutagenesis was performed to change the following residue pairs: S375C/Q1020C, T492C/S1137C, H514C/D1159C, S586C/S1231C, and R613C/A1258C.
X
ABCC1 p.Ser1137Cys 22086917:65:99
status: NEW176 Mutant pairs are S375C/ Q1020C (SQ, black), T492C/S1137C (TS, yellow), H514C/D1159C (HD, pink), S586C/S1231C (SS, green), and R613C/A1258C (RA, cyan).
X
ABCC1 p.Ser1137Cys 22086917:176:50
status: NEW