BIOMOLECULAR SOLUTION X-RAY SCATTERING AT n2STAR BEAMLINE

Year 2022, Volume: 8 Issue: 2, 60 - 69, 30.12.2022

Oktay Göcenler , Cansu Müşerref Yenici , Kerem Kahraman , Cengizhan Büyükdağ , Çağdaş Dağ

https://doi.org/10.22531/muglajsci.1174474

Cited By: 2

Abstract

Small angle X-ray Scattering (SAXS) is a method for determining basic structural characteristics such as the size, shape, and surface of particles. SAXS data can be used to generate low resolution models of biomolecules faster than any other conventional experimental structural biology tool. SAXS data is mostly collected in synchrotron facilities to obtain the best scattering data possible however home source SAXS devices can also generate valuable data when properly optimized. Here, we examined sample data collection and optimization at home source SAXS beamline in terms of the concentration, purity, and duration of data acquisition. We validated that high concentration, monodisperse and ultra pure protein samples obtained by size exclusion chromatography are necessary for generating viable SAXS data using a home source beamline. At least one hour is required to generate a feasible model from SAXS data, although longer data collection times do not always translate to higher resolutions. We show that with small optimizations during data collection and analysis SAXS can characterize properties such as oligomerization, molecular mass, and overall shape of particles in solution under physiological conditions.

Keywords

SAXS, GST, protein, in solution

Supporting Institution

TÜBİTAK

Project Number

120Z594, 221S535, 122Z747

Thanks

We are greatful to Prof. Ziarek at Indiana University-Bloomington for constructive feedback and we apprecıate Ziarek Juniors for their understanding. CD acknowledges support from TÜBİTAK (Project No: 120Z594, 221S535, 122Z747). The authors acknowledge the use of the services and facilities of n2STAR-Koç University Nanofabrication and Nanocharacterization Center for Scientific and Technological Advanced Research.

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