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RESEARCH PRODUCT
ProteoPlex: stability optimization of macromolecular complexes by sparse-matrix screening of chemical space.
Jan-michael PetersGeorg PetzoldJürgen MarklBrenda A. SchulmanHolger StarkVanessa MöllerJuergen OhmerNiels FischerJeremiah J. FryeClemens GrimmKai TittmannAshwin ChariMarc A. JarvisUtz FischerDavid HaselbachOleg M. GanichkinElham PakniaMichael TietzelJan-martin Kirvessubject
Models MolecularProtein FoldingProtein ConformationSupramolecular chemistryBiochemistryArticleProtein structureComputer SimulationMolecular Biologychemistry.chemical_classificationBinding SitesBiomoleculeMolecular biophysicsCell BiologyMolecular machineChemical spacechemistryModels ChemicalChemical physicsMultiprotein ComplexesBiophysicsProtein foldingCrystallizationAlgorithmsSoftwareBiotechnologyMacromoleculeProtein Bindingdescription
Molecular machines or macromolecular complexes are supramolecular assemblies of biomolecules that ensure cellular homeostasis. Structure determination of those complexes in a purified state is often a tedious undertaking due to the compositional complexity and the related relative structural instability. To improve the stability of macromolecular complexes in vitro, we present here a generic method that optimizes the stability, homogeneity and solubility of macromolecular complexes by sparse-matrix screening of their thermal unfolding behaviour in the presence of various buffers and small molecules. The method includes the automated analysis of thermal unfolding curves based on a newly developed biophysical unfolding model for complexes. We found that under stabilizing conditions even large multi-component complexes reveal an almost ideal two-state unfolding behaviour. We envisage an improved biochemical understanding of purified macromolecules as well as a substantial boost in successful macromolecular complex structure determination by both X-ray crystallography and Cryo EM.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2015-09-01 | Nature methods |