Form factor determination of biological molecules with X-ray free electron laser small-angle scattering (XFEL-SAS).
Clement E BlanchetAdam R RoundHaydyn D T MertensKartik AyyerMelissa Ann GraewertSalah AwelDaniel FrankeKaterina DörnerSasa BajtRichard BeanTânia F CustódioRaphael de WijnJuncheng EAlessandra HenkelAndrey GruzinovCy Michael JeffriesYoonhee KimHenry J KirkwoodMarco KloosJuraj KnoškaJayanath C P KoliyaduRomain LetrunChristian LöwJana MakroczyovaAbhishek MallRob MeijersGisel Esperanza Pena MurilloDominik OberthürEkaterina RoundCarolin SeuringMarcin SikorskiPatrik VagovicJoana ValerioTamme WollweberYulong ZhuangJoachim SchulzHeinrich HaasHenry N ChapmanAdrian P MancusoDmitri I SvergunPublished in: Communications biology (2023)
Free-electron lasers (FEL) are revolutionizing X-ray-based structural biology methods. While protein crystallography is already routinely performed at FELs, Small Angle X-ray Scattering (SAXS) studies of biological macromolecules are not as prevalent. SAXS allows the study of the shape and overall structure of proteins and nucleic acids in solution, in a quasi-native environment. In solution, chemical and biophysical parameters that have an influence on the structure and dynamics of molecules can be varied and their effect on conformational changes can be monitored in time-resolved XFEL and SAXS experiments. We report here the collection of scattering form factors of proteins in solution using FEL X-rays. The form factors correspond to the scattering signal of the protein ensemble alone; the scattering contributions from the solvent and the instrument are separately measured and accurately subtracted. The experiment was done using a liquid jet for sample delivery. These results pave the way for time-resolved studies and measurements from dilute samples, capitalizing on the intense and short FEL X-ray pulses.
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