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

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 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 and Pau Bernad Pau Bernad ó Peter Konarev Pau Bernadó Pau Bernad ó ó

Outline Outline Outline Outline 1 1 How to deal with mixtures with ATSAS How to deal with mixtures with ATSAS 1 1 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 Mixture of a few species 3 3 3 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)

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 oligomerization, 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

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

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

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

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

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

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

Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme Thermal unfolding of lysozyme s (1/ Å ) s (1/ s (1/ s (1/Å Å Å) ) )

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

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

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

OLIGOMER OLIGOMER OLIGOMER OLIGOMER in acti in acti in acti in action on on on 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?

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

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

Concentration dependence Concentration dependence Concentration dependence Concentration dependence

OLIGOMER/SASREFMX OLIGOMER/SASREFMX OLIGOMER/SASREFMX OLIGOMER/SASREFMX in acti in acti in acti in action on on on SASREFMX: How does it work? SASREFMX: How does it work? SASREFMX: How does it work? SASREFMX: How does it work?

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

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

Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Folded Unfolded I( s ) ~ 1/ s 4 Scattering I( s ) ~ 1/ s I( s ) ~ 1/ s 2 of a Rod

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

Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Ensemble Optimization Method (EOM) Flexible Rigid

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

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