Acyl-peptide hydrolase and methods of production and use
AssigneeThe General Hospital Corporation
InventorJohn A. Smith
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Abstract
The invention discloses the amino acid sequence of Acyl-Peptide Hydrolase. The invention relates to a DNA molecule encoded for Acyl-Peptide Hydrolase, a vector incorporating the molecule, and the use of such vector to transform a host. The invention further relates to the expression by the host to produce Acyl-Peptide Hydrolase, and the use of APH to catalyze the hydrolysis of an N.sup..alpha. -acetylated peptide or protein or the reaction between an N.sup..alpha. -acetyl amino acid donor and an acceptor protein with a free .alpha.-NH.sub.2 group. The invention further relates to the use of the Acyl-Peptide Hydrolase-encoding sequences in the detection of cancer.
Claims13
A purified nucleic acid molecule which contains a genetic sequence encoding the amino acid sequence shown in FIG. 3.
A purified DNA molecule, wherein said molecule comprises a nucleic acid sequence, said nucleic acid sequence
- 1) encoding a naturally occurring mammalian acyl-peptide hydrolase protein, and
- 2) wherein said nucleotide sequence hybridizes to a cDNA probe complementary to the nucleotide sequence of FIG. 3 or FIG. 4 when the hybridization is performed at about 42.degree. C. in a medium consisting essentially of 50% formamide, 5.times.SSC, 5.times.Denhardt's solution, 10 mM sodium phosphate, 0.1% SDS, 1 mM EDTA and 50 .mu.g/ml of salmon sperm DNA, and wherein washing occurs in a medium consisting essentially of 0.2.times.SSC, 0.1% SDS, 1 mM sodium phosphate, pH 7.0 and 1 mM EDTA at about 55.degree. C.
A method of cloning a DNA molecule that encodes a naturally occurring mammalian acyl-peptide hydrolase protein comprising
- 1) hybridizing a cDNA probe complementary to the nucleotide sequence of FIG. 3 or FIG. 4 to said DNA molecule, wherein the hybridization is performed at about 42.degree. C. in a medium consisting essentially of 50% formamide, 5.times.SSC, 5.times.Denhardt's solution, 10 mM sodium phosphate, 0.1% SDS, 1 mM EDTA and 50 .mu.g/ml sonicated denatured salmon sperm and washing occurs in a medium consisting essentially of 0.2.times.SSC, 0.1% SDS, 1 mM sodium phosphate, pH 7.0 and 1 mM EDTA at 55.degree. C.;
- 2) selecting those DNA molecules that hybridize to said cDNA probe;
- and
- 3) cloning said DNA molecule of part (2).
Dependencies are inferred from the claim text.