Upgrade of macromolecular crystallography beamline BL17U1 at SSRF

Wang, Qisheng; Zhang, Kunhao; Cui, Yin; Wang, Zhijun; Qiangyan, Pan; Liu, Ke; Sun, Binyong; Zhou, Huan; Li, Min-Jun; Xu, Qin; Xu, Chunyan; Yu, Feng; He, Jianhua

doi:10.1007/s41365-018-0398-9

Cited by 318 publications

(253 citation statements)

References 5 publications

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“…Data collection was performed on an FEI Titan Krios equipped with Gatan K2 Summit electron counting camera. Images were recorded in the super-resolution mode using UCSF-Image4 (Wang et al, 2015) at a nominal magnification of 22,500, which corresponds to a final pixel size of 1.306 Å by binning 2 of the original micrographs. For each image stack, a total dose of about 50 electrons per Å 2 at the specimen were equally fractioned into 32 frames with a total exposure time of 8 s. Defocus values ranged from −1.5 to −2.5 μm.…”

Section: Methodsmentioning

confidence: 99%

4.4 Å Resolution Cryo-EM structure of human mTOR Complex 1

et al. 2016

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Mechanistic target of rapamycin (mTOR) complex 1 (mTORC1) integrates signals from growth factors, cellular energy levels, stress and amino acids to control cell growth and proliferation through regulating translation, autophagy and metabolism. Here we determined the cryo-electron microscopy structure of human mTORC1 at 4.4 Å resolution. The mTORC1 comprises a dimer of heterotrimer (mTOR-Raptor-mLST8) mediated by the mTOR protein. The complex adopts a hollow rhomboid shape with 2-fold symmetry. Notably, mTORC1 shows intrinsic conformational dynamics. Within the complex, the conserved N-terminal caspase-like domain of Raptor faces toward the catalytic cavity of the kinase domain of mTOR. Raptor shows no caspase activity and therefore may bind to TOS motif for substrate recognition. Structural analysis indicates that FKBP12-Rapamycin may generate steric hindrance for substrate entry to the catalytic cavity of mTORC1. The structure provides a basis to understand the assembly of mTORC1 and a framework to characterize the regulatory mechanism of mTORC1 pathway.Electronic supplementary materialThe online version of this article (doi:10.1007/s13238-016-0346-6) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

4.4 Å Resolution Cryo-EM structure of human mTOR Complex 1

et al. 2016

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“…Structure Determination-Diffraction data were collected from cryo-cooled crystals at SSRF beamline BL17U1 (51). Data were processed using HKL2000 (45).…”

Section: Methodsmentioning

confidence: 99%

Cyclic AMP Inhibits the Activity and Promotes the Acetylation of Acetyl-CoA Synthetase through Competitive Binding to the ATP/AMP Pocket

Han

Shen

Wang

et al. 2017

Journal of Biological Chemistry

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Edited by Ruma BanerjeeThe high-affinity biosynthetic pathway for converting acetate to acetyl-coenzyme A (acetyl-CoA) is catalyzed by the central metabolic enzyme acetyl-coenzyme A synthetase (Acs), which is finely regulated both at the transcriptional level via cyclic AMP (cAMP)-driven trans-activation and at the post-translational level via acetylation inhibition. In this study, we discovered that cAMP directly binds to Salmonella enterica Acs (SeAcs) and inhibits its activity in a substrate-competitive manner. In addition, cAMP binding increases SeAcs acetylation by simultaneously promoting Pat-dependent acetylation and inhibiting CobB-dependent deacetylation, resulting in enhanced SeAcs inhibition. A crystal structure study and site-directed mutagenesis analyses confirmed that cAMP binds to the ATP/AMP pocket of SeAcs, and restrains SeAcs in an open conformation. The cAMP contact residues are well conserved from prokaryotes to eukaryotes, suggesting a general regulatory mechanism of cAMP on Acs.Acetyl-coenzyme A (acetyl-CoA) is a central metabolic intermediate that modulates the balance of anabolism and catabolism by functioning as the fuel for energy generation or as the precursor of lipogenesis for energy storage in many organisms (1). Acetyl-CoA is also deemed a second messenger for physiological regulation by supplying an acetyl group for protein acetylation (1, 2), an important mechanism of post-translational modification (PTM). Acetyl-coenzyme A synthetase (Acs) 5 (EC 6.2

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“…We would like to thank the staff of GM/CA‐CAT at Advance Photon Source (APS) and the BL17U, BL18U and BL19U1 beamlines at the Shanghai Synchrotron Radiation Facility (SSRF) for their assistance during data collection . YL is supported by the China Postdoctoral Science Foundation (Grant No.…”

Section: Acknowledgementsmentioning

confidence: 99%