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61.
Chapman HN Fromme P Barty A White TA Kirian RA Aquila A Hunter MS Schulz J DePonte DP Weierstall U Doak RB Maia FR Martin AV Schlichting I Lomb L Coppola N Shoeman RL Epp SW Hartmann R Rolles D Rudenko A Foucar L Kimmel N Weidenspointner G Holl P Liang M Barthelmess M Caleman C Boutet S Bogan MJ Krzywinski J Bostedt C Bajt S Gumprecht L Rudek B Erk B Schmidt C Hömke A Reich C Pietschner D Strüder L Hauser G Gorke H Ullrich J Herrmann S Schaller G Schopper F Soltau H Kühnel KU Messerschmidt M 《Nature》2011,470(7332):73-77
X-ray crystallography provides the vast majority of macromolecular structures, but the success of the method relies on growing crystals of sufficient size. In conventional measurements, the necessary increase in X-ray dose to record data from crystals that are too small leads to extensive damage before a diffraction signal can be recorded. It is particularly challenging to obtain large, well-diffracting crystals of membrane proteins, for which fewer than 300 unique structures have been determined despite their importance in all living cells. Here we present a method for structure determination where single-crystal X-ray diffraction 'snapshots' are collected from a fully hydrated stream of nanocrystals using femtosecond pulses from a hard-X-ray free-electron laser, the Linac Coherent Light Source. We prove this concept with nanocrystals of photosystem I, one of the largest membrane protein complexes. More than 3,000,000 diffraction patterns were collected in this study, and a three-dimensional data set was assembled from individual photosystem I nanocrystals (~200?nm to 2?μm in size). We mitigate the problem of radiation damage in crystallography by using pulses briefer than the timescale of most damage processes. This offers a new approach to structure determination of macromolecules that do not yield crystals of sufficient size for studies using conventional radiation sources or are particularly sensitive to radiation damage. 相似文献
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K. R. Henry 《Cellular and molecular life sciences : CMLS》1971,27(10):1191-1191
Zusammenfassung Amino-oxyacetsäure, eine Substanz welche das endogene Gehirnniveau anGABA erhöht, führte in 16 Tage alten Mäusen zu LSD-ähnlichen Verhaltensweisen wie Hypothermie: zu Krämpfen und Rückwärtslaufen. 相似文献
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Endothelins are vascular-derived axonal guidance cues for developing sympathetic neurons 总被引:1,自引:0,他引:1
During development, sympathetic neurons extend axons along a myriad of distinct trajectories, often consisting of arteries, to innervate one of a large variety of distinct final target tissues. Whether or not subsets of neurons within complex sympathetic ganglia are predetermined to innervate select end-organs is unknown. Here we demonstrate in mouse embryos that the endothelin family member Edn3 (ref. 1), acting through the endothelin receptor EdnrA (refs 2, 3), directs extension of axons of a subset of sympathetic neurons from the superior cervical ganglion to a preferred intermediate target, the external carotid artery, which serves as the gateway to select targets, including the salivary glands. These findings establish a previously unknown mechanism of axonal pathfinding involving vascular-derived endothelins, and have broad implications for endothelins as general mediators of axonal growth and guidance in the developing nervous system. Moreover, they suggest a model in which newborn sympathetic neurons distinguish and choose between distinct vascular trajectories to innervate their appropriate end organs. 相似文献
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Bieling P Laan L Schek H Munteanu EL Sandblad L Dogterom M Brunner D Surrey T 《Nature》2007,450(7172):1100-1105
The microtubule cytoskeleton is essential to cell morphogenesis. Growing microtubule plus ends have emerged as dynamic regulatory sites in which specialized proteins, called plus-end-binding proteins (+TIPs), bind and regulate the proper functioning of microtubules. However, the molecular mechanism of plus-end association by +TIPs and their ability to track the growing end are not well understood. Here we report the in vitro reconstitution of a minimal plus-end tracking system consisting of the three fission yeast proteins Mal3, Tip1 and the kinesin Tea2. Using time-lapse total internal reflection fluorescence microscopy, we show that the EB1 homologue Mal3 has an enhanced affinity for growing microtubule end structures as opposed to the microtubule lattice. This allows it to track growing microtubule ends autonomously by an end recognition mechanism. In addition, Mal3 acts as a factor that mediates loading of the processive motor Tea2 and its cargo, the Clip170 homologue Tip1, onto the microtubule lattice. The interaction of all three proteins is required for the selective tracking of growing microtubule plus ends by both Tea2 and Tip1. Our results dissect the collective interactions of the constituents of this plus-end tracking system and show how these interactions lead to the emergence of its dynamic behaviour. We expect that such in vitro reconstitutions will also be essential for the mechanistic dissection of other plus-end tracking systems. 相似文献