1 / 13

Superoxide Dismutase and Amyotrophic Lateral Sclerosis

Superoxide Dismutase and Amyotrophic Lateral Sclerosis. Dan Zollinger PI: Dr. Joe Beckman Linus Pauling Institute. Oxidative Stress and Familial ALS. Motor Neuron Death Progressive Neurodegeneration Muscle Wasting Paralysis Death Mutations to CuZn Superoxide Dismutase (SOD).

yetty
Télécharger la présentation

Superoxide Dismutase and Amyotrophic Lateral Sclerosis

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Superoxide Dismutase and Amyotrophic Lateral Sclerosis Dan Zollinger PI: Dr. Joe Beckman Linus Pauling Institute

  2. Oxidative Stress and Familial ALS • Motor Neuron Death • Progressive Neurodegeneration • Muscle Wasting • Paralysis • Death • Mutations to CuZn Superoxide Dismutase (SOD)

  3. CuZn Superoxide Dismutase Cu2+ + O2- O2 + Cu1+

  4. G85R and ALS • Does G85R bind copper and show superoxide scavenging activity? • Can copper play a role in the disease?

  5. G85R SOD Activity 900 ng 600 ng 300 ng Bruijn, Et. al., Neuron 1997 Nitro Blue Formazan

  6. Mass Spec Data: Recombinant G85R SOD CuZn Metal def. C679H1081N203O224S4CuZn Apo

  7. G85R SOD Crystal Structure Hart, J. Biol. Chem, 2008

  8. Tissue Samples for MS SOD Assay Dorsal Ventral G85R in vivo • Identify metal status of SOD in diseased transgenic mice • Focus on motor neurons in ventral horn of spinal cord

  9. ZipTip® Method

  10. Western Results of Mouse Spinal Cord S P S P Recombinant G85R (300 ng) G85R mouse tissue Non transgenic mouse tissue 5 punches (~250 g/punch) suspended in 25 L of 50mM Ammonium Acetate pH 5.1

  11. G85R Mouse Tissue Apo G85R Apo G85R Metal def. G85R Mouse SOD Metal def. G85R

  12. Toxicity of G85R SOD • Why is G85R more toxic than other fALS linked mutants? • Hypothesis: • G85R SOD as a lower affinity for zinc than other ALS mutants. • Method • Test the metal release of the mutant using metal chelators and compare to other mutants

  13. Joe Beckman Keith Nylin Nathan Lopez Tim Rhoads Kevin Ahern Howard Hughes Medical Institute Biochemistry and Biophysics Department Acknowledgements

More Related