Electronic Resource
Crystal structure of CmABCB1 multi-drug exporter in lipidic mesophase revealed by LCP-SFX
| العنوان: | Crystal structure of CmABCB1 multi-drug exporter in lipidic mesophase revealed by LCP-SFX |
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| المؤلفون: | 50710787, 60452330, 90204487, Pan, Dongqing, Oyama, Ryo, Sato, Tomomi, Nakane, Takanori, Mizunuma, Ryo, Matsuoka, Keita, Joti, Yasumasa, Tono, Kensuke, Nango, Eriko, Iwata, So, Nakatsu, Toru, Kato, Hiroaki |
| بيانات النشر: | International Union of Crystallography (IUCr) 2022-01 |
| نوع الوثيقة: | Electronic Resource |
| مستخلص: | CmABCB1 is a Cyanidioschyzon merolae homolog of human ABCB1, a well known ATP-binding cassette (ABC) transporter responsible for multi-drug resistance in various cancers. Three-dimensional structures of ABCB1 homologs have revealed the snapshots of inward- and outward-facing states of the transporters in action. However, sufficient information to establish the sequential movements of the open–close cycles of the alternating-access model is still lacking. Serial femtosecond crystallography (SFX) using X-ray free-electron lasers has proven its worth in determining novel structures and recording sequential conformational changes of proteins at room temperature, especially for medically important membrane proteins, but it has never been applied to ABC transporters. In this study, 7.7 monoacylglycerol with cholesterol as the host lipid was used and obtained well diffracting microcrystals of the 130 kDa CmABCB1 dimer. Successful SFX experiments were performed by adjusting the viscosity of the crystal suspension of the sponge phase with hydroxypropyl methylcellulose and using the high-viscosity sample injector for data collection at the SACLA beamline. An outward-facing structure of CmABCB1 at a maximum resolution of 2.22 Å is reported, determined by SFX experiments with crystals formed in the lipidic cubic phase (LCP-SFX), which has never been applied to ABC transporters. In the type I crystal, CmABCB1 dimers interact with adjacent molecules via not only the nucleotide-binding domains but also the transmembrane domains (TMDs); such an interaction was not observed in the previous type II crystal. Although most parts of the structure are similar to those in the previous type II structure, the substrate-exit region of the TMD adopts a different configuration in the type I structure. This difference between the two types of structures reflects the flexibility of the substrate-exit region of CmABCB1, which might be essential for the smooth release of various substrates fr |
| مصطلحات الفهرس: | serial crystallography, SFX, protein structures, sample delivery, XFELs, LCP, lipidic mesophase, multi-drug exporters, ABC transporters, Cyanidioschyzon merolae, CmABCB1, journal article |
| URL: | 10.1107/S2052252521011611 |
| الاتاحة: | Open access content. Open access content https://creativecommons.org/licenses/by/4.0 This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
| ملاحظة: | English |
| Other Numbers: | YJ@ oai:repository.kulib.kyoto-u.ac.jp:2433/266705 IUCrJ 9 1 134 145 35059217 2052-2525 1458632946 |
| المصدر المساهم: | KYOTO UNIV From OAIster®, provided by the OCLC Cooperative. |
| رقم الانضمام: | edsoai.on1458632946 |
| قاعدة البيانات: | OAIster |
| الوصف غير متاح. |