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A Comparative Electron Paramagnetic Resonance Study of the Nucleotide-Binding Domains’ Catalytic Cycle in the Assembled Maltose ATP-Binding Cassette Importer

A Comparative Electron Paramagnetic Resonance Study of the Nucleotide-Binding Domains’ Catalytic Cycle in the Assembled Maltose ATP-Binding Cassette Importer

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GROTE, Mathias, Enrica BORDIGNON, Yevhen POLYHACH, Gunnar JESCHKE, Heinz-Jürgen STEINHOFF, Erwin SCHNEIDER, 2008. A Comparative Electron Paramagnetic Resonance Study of the Nucleotide-Binding Domains’ Catalytic Cycle in the Assembled Maltose ATP-Binding Cassette Importer. In: Biophysical Journal. Cell Press. 95(6), pp. 2924-2938. ISSN 0006-3495. eISSN 1542-0086. Available under: doi: 10.1529/biophysj.108.132456

@article{Grote2008-09-15Compa-58680, title={A Comparative Electron Paramagnetic Resonance Study of the Nucleotide-Binding Domains’ Catalytic Cycle in the Assembled Maltose ATP-Binding Cassette Importer}, year={2008}, doi={10.1529/biophysj.108.132456}, number={6}, volume={95}, issn={0006-3495}, journal={Biophysical Journal}, pages={2924--2938}, author={Grote, Mathias and Bordignon, Enrica and Polyhach, Yevhen and Jeschke, Gunnar and Steinhoff, Heinz-Jürgen and Schneider, Erwin} }

Steinhoff, Heinz-Jürgen Polyhach, Yevhen 2008-09-15 2022-09-22T11:49:46Z Schneider, Erwin A Comparative Electron Paramagnetic Resonance Study of the Nucleotide-Binding Domains’ Catalytic Cycle in the Assembled Maltose ATP-Binding Cassette Importer Bordignon, Enrica We present a quantitative analysis of conformational changes of the nucleotide-binding subunits, MalK<sub>2</sub>, of the maltose ATP-binding cassette importer MalFGK2 during the transport cycle. Distance changes occurring between selected residues were monitored in the full transporter by site-directed spin-labeling electron paramagnetic resonance spectroscopy and site-directed chemical cross-linking. We considered S83C and A85C from the conserved Q-loop and V117C located on the outer surface of MalK. Additionally, two native cysteines (C350, C360) were included in the study. On ATP binding, small rearrangements between the native sites, and no distance changes between positions 117 were detected. In contrast, positions 85 come closer together in the ATP-bound state and in the vanadate-trapped intermediate and move back toward the apo-state after ATP hydrolysis. The distance between positions 83 is shown to slightly decrease on ATP binding, and to further decrease after ATP hydrolysis. Results from cross-linking experiments are in agreement with these findings. The data are compared with in silico spin-labeled x-ray structures from both isolated MalK<sub>2</sub> and the MalFGK<sub>2</sub>-E complex. Our results are consistent with a slightly modified “tweezers-like” model of closure and reopening of MalK<sub>2</sub> during the catalytic cycle, and show an unforeseen potential interaction between MalK and the transmembrane subunit MalG. Grote, Mathias Grote, Mathias Jeschke, Gunnar Schneider, Erwin Jeschke, Gunnar eng 2022-09-22T11:49:46Z Polyhach, Yevhen Bordignon, Enrica Steinhoff, Heinz-Jürgen

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