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For those who are interested in details, I will tell the more complete story on Feb 12 here:https://www.nature.com/webcasts/event/using-cryo-em-for-designing-next-gen-therapeutics-against-hiv/ …
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11. Cryo-EM based drug design is real. Still lots of throughput issues ... but we will get there soon ... more microscopes and cryo-EM democratization, ala
@cjrlab and colleagues, will help.Prikaži ovu nitHvala. Twitter će to iskoristiti za poboljšanje vaše vremenske crte. PoništiPoništi -
10. The “substrate envelope” hypothesis (developed by Schiffer lab) is important for drug design. Basically, drugs should occupy molecular envelope of natural substrates. This way, "resistant variants" also compromise fitness and do not persist in clonal populations.
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9. We see many water molecules throughout the active site. Waters are critical to ligand binding. Their displacement can be advantageous to ligand potency. For drug design and computational chemistry, “high-resolution” means seeing waters.
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8. All compounds bind potently to WT intasomes. Only a few bind tightly to viral resistant variants. Drugs need to be designed for targeting the latter. Tackling resistance is the most important problem for antiviral therapy (and for many other therapeutic approaches).
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7. Minor changes to an enzyme active site have profound implications. Drug resistance results from small structural perturbations. See also colleague Peter Cherepanov’s work on an important, patient-derived resistant variant, also out in same issue:https://science.sciencemag.org/content/early/2020/01/30/science.aay4919 …
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6. The substrate really matters! For 10 years, a different retroviral Intasome was used to study INSTIs. However, the same drugs bind differently to HIV intasomes. The latter should be used now for understanding drug resistance and for structure-based designpic.twitter.com/DeEBdhUesH
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5. Sample is far from “ideal”. Pathologically preferred and prone to rapid aggregation, it required 800 mM salt and 6% glycerol in solution. Nonetheless, using tilted data collection, and with cumulative advances in the field, we resolved the active site to ~2.6 A resolutionpic.twitter.com/I08Qda2BSq
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4. We tried other softwares, but for reasons unknown, were not able to reach high-resolution. Datasets will be deposited to
@EMDB_EMPIAR for community to try ... please tell us what we could do better!Prikaži ovu nitHvala. Twitter će to iskoristiti za poboljšanje vaše vremenske crte. PoništiPoništi -
3. Many new advances helped us along the way,
@DTegunov and#WARP. CisTEM for refinement, @TwitterlessTim and@AlexisRohou, advances in Phenix, density modification@TerwillTom, + othersPrikaži ovu nitHvala. Twitter će to iskoristiti za poboljšanje vaše vremenske crte. PoništiPoništi -
2. Initial cryo-EM data was pathologically preferentially oriented. Although we tried different approaches, including novel spotting methods, only tilting the grid solved the problem.
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1. Cryo-EM enables determining structures of HIV integration complexes (intasomes) bound to leading clinically used drugs and pre-clinical drug candidates. One compound (4d) has superior potency to all clinically used drugs against viral resistant variants.
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Taking advantage of the Tweetorial ... I’m excited to share our work, with Dario Passos,
@IlonaJozwik, Min Li,@ForliLab, and@theNCI collaborators pertaining to cryo-EM in drug discovery targeting HIV integration, out now: https://science.sciencemag.org/content/early/2020/01/29/science.aay8015 …. We make the following advances:Prikaži ovu nitHvala. Twitter će to iskoristiti za poboljšanje vaše vremenske crte. PoništiPoništi -
Finally got a chance to sit down and read this. Great stuff
@mcianfro! Love the incorporation of tilts ... images even look tilted in figures (Fig 2A,C)
. What are the challenges for extending to 3D classification?https://twitter.com/mcianfro/status/1208450336274432004 …
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This is a neat application of AFM to clarify an important question in chromatin biology. When is this sample going onto cryo grids?
https://www.biorxiv.org/content/10.1101/2019.12.13.867358v1 …Hvala. Twitter će to iskoristiti za poboljšanje vaše vremenske crte. PoništiPoništi -
Is 50 kDa now becoming “routine”?!
#CryoEM#NewStandardshttps://twitter.com/tanyongzi/status/1205147620366262272 …Hvala. Twitter će to iskoristiti za poboljšanje vaše vremenske crte. PoništiPoništi -
New advances from colleagues across the street, continuing to break Cryo-EM barriershttps://twitter.com/markherzik/status/1199356822039064577 …
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Reminds me of the famous saying by Frederick Douglass: “It is easier to build strong children than to repair broken men” ... spreading the word to allies (and passive opposition)https://ensia.com/voices/climate-change-deniers/ …
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From Pedal the Cause
@PedalSD yesterday @PetcoPark. $2.1 million raised, still three weeks left. 100% of funds go to research. Please spread the word.#endcancerpic.twitter.com/wGQd3o59Jo
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And the link to the original article in case you haven’t read it yet.https://journals.iucr.org/m/issues/2019/06/00/eh5004/ …
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