Elan Pharmaceuticals, Inc.

Pharmacia & Upjohn Company

Pfizer Corporation

Thomas L Emmons

Alfredo Giuseppe Tomasselli

Robert L Heinrikson

Mark E Gurney

Michael Jerome Bienkowski

Riqiang Yan

Ana Mildner

Donna Paddock

Timothy Earl Benson

Jim D Durbin

The present invention is directed to novel substrates for Hu-Asp. More particularly, the invention provides peptide substrates and fusion polypeptide substrates comprising a &#x3b2;-secretase cleavage site. Methods and compositions for making and using the peptides are disclosed.

Substrates and assays for &#x3b2;-secretase

The invention provides methods for efficient recombinant expression, refolding, and purification of Beta-site APP cleaving enzyme (BACE) polypeptides. In various aspects, the method includes the steps of expressing a recombinant construct in bacteria, dissolving inclusion bodies with a denaturant at high pH in the presence of a reducing agent, diluting the solubilized BACE polypeptide in an aqueous solution at a temperature of about 1&#xb0; C. to 15&#xb0; C., and incubating the diluted sample at a temperature of about 4&#xb0; C. to 15&#xb0; C. until the recombinant BACE polypeptide folds into an active enzyme.

Method for refolding enzymes

The x-ray crystal structure of human BACE or BACE-like proteins is useful for solving the structure of other molecules or molecular complexes, and identifying and/or designing potential modifiers of human BACE activity.

Crystal of HIV protease-cleaved human beta secretase and method for crystallization thereof

Substrates and assays for &#x3b2;-secretase activity

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Wentzville, MO, United States of America

Thomas L Emmons