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Hot off the press excitement for #structuralbiology combining NanoTemper Technologies label-free thermal unfolding #nanoDSF with #CryoEM! 🎆 A global team of researchers from Stockholm University, Harvard University, Boston Children's Hospital and Ipsen Bioinnovation just published this exciting work in Nature Portfolio's Communication Chemistry investigating a new botulinum neurotoxin serotype, BoNT/X, with promising therapeutic properties, and how the presence or absence of it's protective partner protein (NTNH) that stabilized the BoNT/X affects potency. Read more about this exciting #research here: https://lnkd.in/g9rfPNv8 #biophysics #protein #toxin #therapeutic #teamwork
Measuring time at the CryoEM is extremely valuable and expensive. Make sure that you only examine samples that are well prepared.
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2moIan Stanton, Ph.D. Thanks for sharing. This was a great read. Reading the paper, I was curious as to why a pH of 5.5 was employed for the preparation of the sample for #CryoEM analysis of the BoNT-NTNH complex. It turns out that: 1. The BoNT-NTNH complexes are held together by several pH-dependent contacts 2. The role of the partner protein NTNH is to stabilize the Botulinum neurotoxins (BoNTs) and resist the acidic and proteolytic attacks 3. SEC showed that the BoNT/X and NTNH/X is stable at pH 5.5 (falls apart at pH 9.5) 4. pH-dependent trypsin proteolysis also supported that at pH 5.5 BoNT/X-NTNH/X complex is largely resistant to trypsin 5. Lastly #nanoDSF also confirmed that the complex is very stable at pH 5.5 :)