Crystals of Cry11Aa were centrifuged for 5 min at 5000 × g during the buffer wash and washed twice with ammonium acetate buffer (pH adjusted to 6.4 with acetic acid). Pelleted crystals were then dissolved in ammonium acetate buffer (pH adjusted to 11.5 using ammonium hydroxide). Gold-coated capillary emitters were prepared as previously described and used to load the protein sample105 (link). The sample was analyzed on a Synapt G1 mass spectrometer (Waters Corporation). The instrument was tuned to preserve non-covalent interactions. Briefly, the capillary voltage was set to 1.60 kV, the sampling cone voltage was 20 V, the extraction cone voltage was 5 V, the source temperature was 80 °C, the trap transfer collision energy was 10 V, and the trap collision energy (CE) was set at 30 V. For MS/MS characterization, a particular charge state was isolated in the quadrupole and the complex was dissociated by application of 200 V of CE. The data collected were deconvoluted and analyzed using UniDec106 (link).
Tetreau G., Sawaya M.R., De Zitter E., Andreeva E.A., Banneville A.S., Schibrowsky N.A., Coquelle N., Brewster A.S., Grünbein M.L., Kovacs G.N., Hunter M.S., Kloos M., Sierra R.G., Schiro G., Qiao P., Stricker M., Bideshi D., Young I.D., Zala N., Engilberge S., Gorel A., Signor L., Teulon J.M., Hilpert M., Foucar L., Bielecki J., Bean R., de Wijn R., Sato T., Kirkwood H., Letrun R., Batyuk A., Snigireva I., Fenel D., Schubert R., Canfield E.J., Alba M.M., Laporte F., Després L., Bacia M., Roux A., Chapelle C., Riobé F., Maury O., Ling W.L., Boutet S., Mancuso A., Gutsche I., Girard E., Barends T.R., Pellequer J.L., Park H.W., Laganowsky A.D., Rodriguez J., Burghammer M., Shoeman R.L., Doak R.B., Weik M., Sauter N.K., Federici B., Cascio D., Schlichting I, & Colletier J.P. (2022). De novo determination of mosquitocidal Cry11Aa and Cry11Ba structures from naturally-occurring nanocrystals. Nature Communications, 13, 4376.
Other organizations :
Institut de Biologie Structurale, Centre National de la Recherche Scientifique, CEA Grenoble, Commissariat à l'Énergie Atomique et aux Énergies Alternatives, Université Grenoble Alpes, University of California, Los Angeles, Institut Laue-Langevin, Lawrence Berkeley National Laboratory, Max Planck Institute for Medical Research, Linac Coherent Light Source, SLAC National Accelerator Laboratory, Texas A&M University, University of California, Riverside, European X-Ray Free-Electron Laser, European Synchrotron Radiation Facility, University of Southern California, Laboratoire d'Écologie Alpine, Université Claude Bernard Lyon 1, École Normale Supérieure de Lyon
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to
get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
