Protein unfolding and flexible systems Protein unfolding and - - PowerPoint PPT Presentation

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Protein unfolding and flexible systems Protein unfolding and - - PowerPoint PPT Presentation

Mar 04, 2023 307 likes •575 views

European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation BioSAXS group

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Protein unfolding and flexible systems Protein unfolding and flexible systems Protein unfolding and flexible systems Protein unfolding and flexible systems

  • Dr. Weifeng Shang
  • Dr. Weifeng Shang
  • Dr. Weifeng Shang
  • Dr. Weifeng Shang 尚伟峰

Acknowledgement: Peter Konarev Peter Konarev Peter Konarev Peter Konarev and Pau Bernad Pau Bernad Pau Bernad Pau Bernadó ó ó ó

European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation European Molecular Biology Laboratory Hamburg Outstation BioSAXS group BioSAXS group BioSAXS group BioSAXS group

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Outline Outline Outline Outline 1 1 1 1 How to deal with mixtures with ATSAS How to deal with mixtures with ATSAS How to deal with mixtures with ATSAS How to deal with mixtures with ATSAS 2 Protein unfolding 2 Protein unfolding 2 Protein unfolding 2 Protein unfolding 3 3 3 3 Mixture of a few species Mixture of a few species Mixture of a few species Mixture of a few species 4 4 4 4 Flexible systems Flexible systems Flexible systems Flexible systems 5 Ensemble Optimization Method (EOM) 5 Ensemble Optimization Method (EOM) 5 Ensemble Optimization Method (EOM) 5 Ensemble Optimization Method (EOM)

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Mixtures with ATSAS Mixtures with ATSAS Mixtures with ATSAS Mixtures with ATSAS A few known species A few known species A few known species A few known species

  • ligomerization, dissociation of complex/oligomer

OLIGOMER/SASREFMX OLIGOMER/SASREFMX OLIGOMER/SASREFMX OLIGOMER/SASREFMX Unknown species Unknown species Unknown species Unknown species SVDPLOT/MIXTURE SVDPLOT/MIXTURE SVDPLOT/MIXTURE SVDPLOT/MIXTURE Intrinsically unfolded proteins and multidomain Intrinsically unfolded proteins and multidomain Intrinsically unfolded proteins and multidomain Intrinsically unfolded proteins and multidomain proteins with flexible linkers proteins with flexible linkers proteins with flexible linkers proteins with flexible linkers Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ranch + Gajoe Ranch + Gajoe Ranch + Gajoe Ranch + Gajoe

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Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme

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Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme

10 mg/ml lysozyme in 40 mM NaAc, 150 mM NaCl 10 mg/ml lysozyme in 40 mM NaAc, 150 mM NaCl 10 mg/ml lysozyme in 40 mM NaAc, 150 mM NaCl 10 mg/ml lysozyme in 40 mM NaAc, 150 mM NaCl 10 mg/ml lysozyme in water 10 mg/ml lysozyme in water 10 mg/ml lysozyme in water 10 mg/ml lysozyme in water

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Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme

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Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme

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Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme

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Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme

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Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme

s (1/ s (1/ s (1/ s (1/Å Å Å Å) ) ) )

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Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme

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Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme

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mammalian lipoxygenase mammalian lipoxygenase mammalian lipoxygenase mammalian lipoxygenase rabbit lipoxygenase 2p0m rabbit lipoxygenase 2p0m rabbit lipoxygenase 2p0m rabbit lipoxygenase 2p0m

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OLIGOMER OLIGOMER OLIGOMER OLIGOMER in acti in acti in acti in action

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Identification of the tetramer in P222 Identification of the tetramer in P222 Identification of the tetramer in P222 Identification of the tetramer in P222 Problem! How to explain? Problem! How to explain? Problem! How to explain? Problem! How to explain?

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OLIGOMER OLIGOMER OLIGOMER OLIGOMER in acti in acti in acti in action

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Flexible systems + Mixtures Flexible systems + Mixtures Flexible systems + Mixtures Flexible systems + Mixtures

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Concentration dependence Concentration dependence Concentration dependence Concentration dependence

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OLIGOMER/SASREFMX OLIGOMER/SASREFMX OLIGOMER/SASREFMX OLIGOMER/SASREFMX in acti in acti in acti in action

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SASREFMX: How does it work? SASREFMX: How does it work? SASREFMX: How does it work? SASREFMX: How does it work?

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SASREFMX SASREFMX SASREFMX SASREFMX in acti in acti in acti in action

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Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM)

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Folded Unfolded I(s) ~ 1/s4 I(s) ~ 1/s I(s) ~ 1/s2

Scattering

  • f a Rod

Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM)

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Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM)

SAXS as a tool to validate structural models with flexibility Detection of flexibility in SAXS data Ensemble methods to characterize flexible biomolecules

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Flexible Rigid

Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM)

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EOM on titin EOM on titin EOM on titin EOM on titin SAXS data on titin constructs: SAXS data on titin constructs: SAXS data on titin constructs: SAXS data on titin constructs: A164-A169 (5 Ig domains) A164-A169 (5 Ig domains) A164-A169 (5 Ig domains) A164-A169 (5 Ig domains) A168-A169-A170-TK-M1-M2 (5 A168-A169-A170-TK-M1-M2 (5 A168-A169-A170-TK-M1-M2 (5 A168-A169-A170-TK-M1-M2 (5 domains) domains) domains) domains) A170-TK A170-TK A170-TK A170-TK

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EOM in action EOM in action EOM in action EOM in action