Back to results
US 9,290,757 B2Granted
Protease screening methods and proteases identified thereby
AssigneeCatalyst Biosciences, Inc.
InventorEdwin L. Madison
Life of this patent
Estimating the term…
Abstract
Methods for identifying modified proteases with modified substrate specificity or other properties are provided. The methods screen candidate and modified proteases by contacting them with a substrate, such as a serpin, an alpha macroglobulins or a p35 family protein or modified serpins and modified p35 family members or modified alpha macroglobulins, that, upon cleavage of the substrate, traps the protease by forming a stable complex. Also provided are modified proteases.
Claims12
A modified membrane type serine protease 1 (MT-SP1) polypeptide or a catalytically active portion thereof, comprising:
- a) an amino acid substitution in an unmodified MT-SP1 at a position corresponding to the position 60(g), based on chymotrypsin numbering, whereby substrate specificity or activity is altered compared to the MT-SP1 polypeptide not containing the amino acid substitution;
- or
- b) an amino acid substitution at a position corresponding to T98 or F99 based on chymotrypsin numbering, whereby substrate specificity or activity is altered compared to the MT-SP1 polypeptide not containing the substitution, wherein the substitution is a T98P or a F99L substitution,
- wherein the unmodified MT-SP1 polypeptide or catalytically active fragment is selected from the group consisting of:
- an MT-SP1 polypeptide having the sequence of amino acids set forth in SEQ ID NO:253 or 515, or a sequence of amino acids that exhibits at least 95% sequence identity to SEQ IDNO:253 or 515;
- and
- a catalytically active portion of an MT-SP1 having the sequence of amino acids set forth in SEQ ID NO:
- 505 or 507 or a sequence of amino acids that exhibits at least 95% sequence identity to SEQ ID NO:505 or 507.
5 more elements
Dependencies are inferred from the claim text.