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US 5,932,433 AGranted
Biotinylation of proteins
AssigneeAffymax Technologies N.V.
InventorPeter J. Schatz
Life of this patent
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Abstract
Biotinylation peptides can be fused to other peptides or proteins of interest using recombinant DNA techniques to provide efficient methods for biotinylating the resulting fusion proteins in vivo or in vitro.
Claims6
A method for identifying a biotinylating enzyme, said method comprising:
- (a) on a surface of a substrate, providing a fusion protein comprising a recombinant protein and a peptide that comprises an amino acid sequence defined by;
- Leu Xaa.sub.1 Xaa.sub.2 Ile Xaa.sub.3 Xaa.sub.4 Xaa.sub.5 Xaa.sub.6 Lys Xaa.sub.7 Xaa.sub.8 Xaa.sub.9 Xaa.sub.10 (SEQ ID NO:1), where Xaa.sub.1 is any amino acid;
- Xaa.sub.2 is any amino acid other than Leu, Val, Ile, Trp, Phe, or Tyr;
- Xaa.sub.3 is Phe or Leu;
- Xaa.sub.4 is Glu or Asp;
- Xaa.sub.5 is Ala, Gly, Ser, or Thr;
- Xaa.sub.6 is Gln or Met;
- Xaa.sub.7 is Ile, Met, or Val;
- Xaa.sub.8 is Glu, Leu, Val, Tyr, or Ile;
- Xaa.sub.9 is Trp, Tyr, Val, Phe, Leu, or Ile;
- and Xaa.sub.10 is any amino acid other than Asp or Glu, wherein said peptide is capable of being biotinylated by a biotin ligase at said lysine residue adjacent to Xaa.sub.6 ;
- and is 13 to 50 amino acids in length;
- (b) in a predefined region of the surface of the substrate, contacting said fusion protein with an enzyme;
- and
- (c) determining whether the fusion protein has been biotinylated.
12 more elements
Dependencies are inferred from the claim text.