Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Reexamination Certificate
2001-06-13
2004-05-11
Prouty, Rebecca (Department: 1652)
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
C435S023000, C435S069100, C435S325000, C435S252300, C435S320100, C435S219000, C536S023200, C536S024300, C536S024310, C536S023100
Reexamination Certificate
active
06734006
ABSTRACT:
TECHNICAL FIELD OF THE INVENTION
The invention relates to the area of regulation human transmembrane serine protease activity to provide therapeutic effects.
BACKGROUND OF THE INVENTION
Metastasizing cancer cells invade the extracellular matrix using plasma membrane protrusions that contact and dissolve the matrix with proteases. Agents that inhibit such protease activity can be used to suppress metastases. Proteases also are expressed during development, when degradation of the extracellular matrix is desired. In cases where appropriate extracellular matrix degradation does not occur, supplying a molecule with a protease activity can provide the necessary enzymatic activity. Thus, there is a need in the art for identifying new proteases and methods of regulating extracellular matrix degradation.
SUMMARY OF THE INVENTION
It is an object of the invention to provide reagents and methods of regulating human transmembrane serine protease. These and other objects of the invention are provided by one or more of the embodiments described below.
One embodiment of the invention is a cDNA encoding a polypeptide comprising an amino acid sequence selected from the group consisting of (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), and (c) biologically active variants thereof.
Yet another embodiment of the invention is an expression vector comprising a polynucleotide which encodes a polypeptide comprising an amino acid sequence selected from the group consisting of (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), and (c) biologically active variants thereof.
Another embodiment of the invention is a host cell comprising an expression vector which encodes a polypeptide comprising an amino acid sequence selected from the group consisting of (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), and (c) biologically active variants thereof.
Still another embodiment of the invention is a purified polypeptide comprising an ammo acid sequence selected from the group consisting of (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), and (c) biologically active variants thereof.
Even another embodiment of the invention is a fusion protein comprising a polypeptide consisting of an amino acid sequence selected from the group consisting of (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), and (c) biologically active variants thereof.
Another embodiment of the invention is a method of producing a polypeptide comprising an amino acid sequence selected from the group consisting of (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), and (c) biologically active variants thereof. A host cell comprising an expression vector that encodes the polypeptide is cultured under conditions whereby the polypeptide is expressed. The polypeptide is isolated.
Yet another embodiment of the invention is a method of detecting a coding sequence for a polypeptide comprising an amino acid sequence selected from the group consisting of (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), and (c) biologically active variants thereof. A polynucleotide comprising 11 contiguous nucleotides selected from the group consisting of (a) the complement of the nucleotide sequence shown in SEQ ID NO:11, (b) the complement of the coding sequence of the cDNA insert of plasmid pCRII-TMSP3, (c) a polynucleotide that hybridizes under stringent conditions to (a) or (b), (d) a polynucleotide having a nucleic acid sequence that deviates from the nucleic acid sequences specified in (a) to (c) due to the degeneration of the genetic code, and (e) a polynucleotide that represents a fragment, derivative, or allelic variation of a nucleic acid sequence specified in (a) to (d) is hybridized to nucleic acid material of a biological sample to form a hybridization complex. The hybridization complex is detected.
Even another embodiment of the invention is a kit far detecting a coding sequence for a polypeptide comprising an amino acid sequence selected from the group consisting of (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), and (c) biologically active variants thereof. The kit comprises a polynucleotide and instructions for detecting the coding sequence. The polynucleotide comprises 11 contiguous nucleotides selected from the group consisting of (a) the complement of the nucleotide sequence shown in SEQ ID NO:11, (b) the complement of the coding sequence of the cDNA insert of plasmid pCRII-TMSP3 to nucleic acid material of a biological sample to form a hybridization complex, (c) a polynucleotide that hybridizes under stringent conditions to (a) or (b), (d) a polynucleotide having a nucleic acid sequence that deviates from the nucleic acid sequences specified in (a) to (c) due to the degeneration of the genetic code, and (e) a polynucleotide that represents a fragment, derivative, or allelic variation of a nucleic acid sequence specified in (a) to (d).
Still another embodiment of the invention is a method of detecting a polypeptide comprising an amino acid sequence selected from the group consisting of (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), and (c) biologically active variants thereof. A biological sample is contacted with a reagent that specifically binds to the polypeptide to form a reagent-polypeptide complex. The reagent-polypeptide complex is detected.
Yet another embodiment of the invention is a kit for detecting a polypeptide comprising an amino acid sequence selected from the group consisting of (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), and (c) biologically active variants thereof. The kit comprises an antibody which specifically binds to the polypeptide and instructions for detecting the polypeptide.
Even another embodiment of the invention is a method of screening for agents that can regulate an activity of a human transmembrane serine protease. A test compound is contacted with a polypeptide comprising an amino acid sequence selected from the group consisting of: (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), and (c) biologically active variants thereof. Binding of the test compound to the polypeptide is detected. A test compound that binds to the polypeptide is thereby identified as a potential agent for regulating the activity of the human transmembrane serine protease.
A further embodiment of the invention is a method of screening for therapeutic agents that can regulate an enzymatic activity of a human transmembrane serine protease. A test compound is contacted with a polypeptide comprising an amino acid sequence selected from the group consisting of: (a) the amino acid sequence shown in SEQ ID NO:12, (b) the amino acid sequence encoded by a cDNA insert contained within plasmid pCRII-TMSP3 (ATCC Accession No. PTA-3433), end (c) biologically active variants thereof. The enzymatic activity of the polypeptide is d
Gedrich Richard W.
Xiao Yonghong
Banner & Witcoff , Ltd.
Bayer Aktiengesellschaft
Prouty Rebecca
Ramirez Delia
LandOfFree
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