Molecules and methods for iterative polypeptide analysis and processing
AssigneeWashington University
InventorsJames J. Havranek, Benjamin Borgo
Life of this patent
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Abstract
Reagents and methods for the digital analysis of proteins or peptides are provided. Specifically provided herein are proteins for identifying the N-terminal amino acid or N-terminal phosphorylated amino acid of a polypeptide. Also, an enzyme for use in the cleavage step of the Edman degradation reaction and a method for using this enzyme are described.
Claims32
1-83. (canceled)
An isolated N-terminal amino acid binding protein (NAAB), comprising a modified, non-naturally occurring tRNA synthetase (RS) that selectively binds to a N-terminal amino acid residue of a polypeptide with at least about a 1.5:1 ratio of specific to non-specific binding.
A method for making a N-terminal amino acid binding (NAAB) protein that selectively binds to a N-terminal amino acid residue of a polypeptide, the method comprising:
- identifying an amino acid binding domain of a tRNA synthetase (RS);
- introducing one or more mutations into the amino acid binding domain to form a NAAB;
- and optionally
- assaying the NAAB for specific binding to the N-terminal amino acid residue of a polypeptide.
A kit for analyzing or sequencing a polypeptide comprising:
- one or more N-terminal amino acid binding proteins (NAABs), wherein each of the one or more NAABs selectively binds to a N-terminal amino acid residue of a polypeptide;
- an Edman degradation enzyme;
- and
- instructions for using the NAABs and the Edman degradation enzyme for analyzing or sequencing a polypeptide.
Dependencies are inferred from the claim text.