Parallel screening and optimization of protein constructs for structural studies

Rasia, Rodolfo M.; Noirclerc‐Savoye, Marjolaine; Bologna, Nicolas Gerardo; Gallet, Benoît; Plevin, Michael J.; Blanchard, Laurence; Palatnik, Javier F.; Brutscher, Bernhard; Vernet, Thierry; Boisbouvier, Jérôme

doi:10.1002/pro.46

Cited by 8 publications

(1 citation statement)

References 26 publications

(28 reference statements)

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“…As for all structural techniques (including NMR, mass spectrometry and crystallography), sample quality plays a pivotal role in the success of a single particle analysis (SPA) cryoEM campaign and the availability of consistently reliable samples is essential in establishing a robust platform for routine determination of 3D structures [35][36][37][38] .…”

Section: Resultsmentioning

confidence: 99%

The CryoEM structure of human serum albumin in complex with ligands

Catalano,

Lucier,

et al. 2024

Preprint

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Human serum albumin (HSA) is the most prevalent plasma protein in the human body, accounting for 60% of the total plasma protein. HSA plays a major pharmacokinetic function, serving as a facilitator in the distribution of endobiotics and xenobiotics within the organism. In this paper we report the cryoEM structures of HSA in the apo form and in complex with two ligands (salicylic acid and teniposide) at a resolution of 3.5, 3.7 and 3.4 Å, respectively. We expand upon previously published work and further demonstrate that sub-4 Å maps of ∼60 kDa proteins can be routinely obtained using a 200 kV microscope, employing standard workflows. Most importantly, these maps allowed for the identification of small molecule ligands, emphasizing the practical applicability of this methodology and providing a starting point for subsequent computational modeling and in silico optimization.

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Section: Resultsmentioning

confidence: 99%

The CryoEM structure of human serum albumin in complex with ligands

Catalano,

Lucier,

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

References

2012

NMR of Biomolecules

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Selective Isotopic Unlabeling of Proteins Using Metabolic Precursors: Application to NMR Assignment of Intrinsically Disordered Proteins

2012

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Selective isotopic unlabeling of proteins can provide important residue-type information as well as reduce congestion of NMR spectra. However, metabolic scrambling often complicates the final isotope-labeling pattern. Here, an array of metabolic precursors is used to perform robust, residue-specific unlabeling of proteins. The resulting isotopic-labeling patterns are predictable and nicely complement NMR experiments that differentiate residue types. This approach has widespread applications, but it is particularly relevant for proteins that lack sequence complexity or a defined tertiary structure.

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Parallel screening and optimization of protein constructs for structural studies

Cited by 8 publications

References 26 publications

The CryoEM structure of human serum albumin in complex with ligands

The CryoEM structure of human serum albumin in complex with ligands

References

Selective Isotopic Unlabeling of Proteins Using Metabolic Precursors: Application to NMR Assignment of Intrinsically Disordered Proteins

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