the effect of copper chelators on zn(-) superoxide dismutase
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The effect of copper chelators on Zn(-) Superoxide Dismutase. Lucy Brennan Dr. Joe Beckman Dr. Kari Trumbull. Amyotrophic Lateral Sclerosis. Motor neuron disease characterized by progressive paralysis 10% of cases are familial Mutations in SOD account for up to 7% of fALS. - PowerPoint PPT PresentationTRANSCRIPT
The effect of copper chelators on Zn(-) Superoxide DismutaseThe effect of copper chelators on Zn(-) Superoxide Dismutase
Lucy Brennan
Dr. Joe Beckman
Dr. Kari Trumbull
Lucy Brennan
Dr. Joe Beckman
Dr. Kari Trumbull
Amyotrophic Lateral SclerosisAmyotrophic Lateral Sclerosis
Motor neuron disease characterized by progressive paralysis
10% of cases are familial Mutations in SOD account for up to 7% of
fALS
Motor neuron disease characterized by progressive paralysis
10% of cases are familial Mutations in SOD account for up to 7% of
fALS
Superoxide dismutaseSuperoxide dismutase
(Cu2+ Zn)SOD+O2---> (Cu1+ Zn)SOD+O2
(Cu1+ Zn)SOD +O2-+2H+--> (Cu2+ Zn)SOD+H2O2
Mutations linked to structural instability Zn(-)SOD has a toxic gain of function believed to
underlie fALS (Cu1+)SOD +O2--> (Cu2+)SOD+ 02
-
(Cu2+ Zn)SOD+O2---> (Cu1+ Zn)SOD+O2
(Cu1+ Zn)SOD +O2-+2H+--> (Cu2+ Zn)SOD+H2O2
Mutations linked to structural instability Zn(-)SOD has a toxic gain of function believed to
underlie fALS (Cu1+)SOD +O2--> (Cu2+)SOD+ 02
-
Copper chelatorsCopper chelators
Dimercaptrol (BAL)
treatment for heavy metal toxicity and Wilson’s disease
Sulfhydral complex
Dimercaptrol (BAL)
treatment for heavy metal toxicity and Wilson’s disease
Sulfhydral complex
GoalsGoals
Validation of WST assay To find a Cu-chelator that effectively
removes copper from SOD Modifying the Cu-chelator into a
potential drug for ALS
Validation of WST assay To find a Cu-chelator that effectively
removes copper from SOD Modifying the Cu-chelator into a
potential drug for ALS
WST assayWST assay
Superoxide detectionSuperoxide detection
An assay including potassium superoxide and WST-1 was performed
Addition of KO2- was
added to the assay WST has been
reduced
An assay including potassium superoxide and WST-1 was performed
Addition of KO2- was
added to the assay WST has been
reduced
Xanthine assayXanthine assay
WST-1 + Xanthine and Xanthine oxidase An increased
absorbance was noted, indicating that WST-1 is reacting with superoxide
WST-1 + Xanthine and Xanthine oxidase An increased
absorbance was noted, indicating that WST-1 is reacting with superoxide
Addition of BAL with Zn(-)SODAddition of BAL with Zn(-)SOD
ImplicationsImplications
Validation of WST assay Limitations
Insight into activity of Zn(-)SOD Possible chelation therapy for ALS
Viable test for such compounds
Validation of WST assay Limitations
Insight into activity of Zn(-)SOD Possible chelation therapy for ALS
Viable test for such compounds
AcknowledgementsAcknowledgements
HHMI Kevin Ahern Dr. Joe Beckman Dr. Kari Trumbull
HHMI Kevin Ahern Dr. Joe Beckman Dr. Kari Trumbull