secretase as a drug target in Alzheimers disease

1
?-secretase as a drug target in Alzheimers
disease

• Leading cause of dementia
• Amyloid plaques
• Cleavage of APP by by ?-secretase and ?-secretase

2

• Inhibition of either ?-or ?-secretase decreases
  A? production
• For the search for potential inhibitors there is
  a need for sufficient amounts of expressed and
  active BACE

3

• The first goal with this project is to mutate
  BACE to make it possible to express soluble BACE
  in E. coli with correct folding and with high
  activity.
• The second goal is to find a BACE inhibitor with
  suitable pharmacological properties.

4
Structural information

• BACE is a membrane-associated aspartic protease.
• The conserved residues and active site residues
  Asp32 and Asp228 are located in the center of the
  cleft in the bilobal structure

5
Cellular mechanisms
6
Inhibition of BACE

• Peptidomimetic inhibitors and nonpeptidomimetic
  inhibitors

7
Experimental Plan
8
Site directed mutagenesis

• Download the structure
• Site directed mutagenesis
• Transformation
• Expression
• Purification
• Random mutagenesis

9

• Assay
• Inhibitor studies