AMID protein, nucleic acid molecules, and uses thereof
OwnerHong-Bing Shu, Liang-Guo Xu +1inventors
InventorsHong-Bing Shu, Liang-Guo Xu, Min Wu
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Abstract
Described here is the cloning and characterization of a novel AIF homologous molecule designated AMID, which is co-localized with mitochondria, and which induces cell death with characteristic apoptotic morphology, independent of caspase activation and p53. Bcl2. Disclosed herein are AMID proteins, fragments and homologues, as well as nucleic acid molecules encoding such proteins, and methods of making and using the same.
Claims46
An isolated protein comprising an amino acid sequence selected from the group consisting of:
- a) an amino acid sequence comprising SEQ ID NO:2;
- b) a biologically active fragment of SEQ ID NO:2;
- and
- c) an amino acid sequence that is at least about 50% identical to SEQ ID NO:2, wherein said amino acid sequence has a biological activity of SEQ ID NO:2.
An isolated fusion protein comprising:
- a) a first amino acid sequence selected from the group consisting of:
- i) an amino acid sequence comprising SEQ ID NO:2;
- ii) a biologically active fragment of SEQ ID NO:2;
- and
- iii) an amino acid sequence that is at least about 50% identical to SEQ ID NO:2 and has a biological activity of SEQ ID NO:2;
- and
- b) a second amino acid sequence that is heterologous to said first amino acid sequence, wherein said first and second are linked to form a fusion protein.
An isolated antibody or antigen binding fragment thereof that selectively binds to SEQ ID NO:2.
A composition comprising at least about 1 μg of an isolated protein selected from the group consisting of:
- a) an amino acid sequence comprising SEQ ID NO:2;
- b) a biologically active fragment of SEQ ID NO:2;
- and
- c) an amino acid sequence that is at least about 50% identical to SEQ ID NO:2, and has a biological activity of SEQ ID NO:2.
An isolated nucleic acid molecule consisting essentially of a nucleic acid sequence selected from the group consisting of:
- a) a nucleic acid sequence encoding SEQ ID NO:2;
- b) a nucleic acid sequence encoding a biologically active fragment of SEQ ID NO:2;
- c) a nucleic acid sequence encoding an amino acid sequence that is at least about 50% identical to SEQ ID NO:2 and has a biological activity of SEQ ID NO:2;
- and
- d) a nucleic acid sequence that is fully complementary to any of the nucleic acid sequences of (a)-(c).
An oligonucleotide consisting essentially of at least 22 consecutive nucleotides of SEQ ID NO:
- 1 or SEQ ID NO:5, or the complement thereof.
A method for detecting the presence of a cancer in a patient, comprising the steps of:
- a) obtaining a biological sample comprising cells from a patient;
- b) detecting in said cells the expression of a protein comprising SEQ ID NO:2 or a nucleic acid molecule comprising SEQ ID NO:5;
- and
- c) comparing the level of expression of the protein or nucleic acid molecule detected in (b) to a control expression level as an indicator of the presence of a cancer in the patient, wherein a decrease in the expression of said protein or said nucleic acid molecule as compared to the control is an indicator of a positive diagnosis of cancer in said patient.
A method for inducing apoptosis in a cell, comprising contacting a cell intracellularly with a compound selected from the group consisting of:
- a) an isolated protein comprising an amino acid sequence selected from the group consisting of:
- i) an amino acid sequence comprising SEQ ID NO:2;
- ii) a biologically active fragment of SEQ ID NO:2;
- and
- iii) an amino acid sequence that is at least about 50% identical to SEQ ID NO:2, wherein said amino acid sequence has a biological activity of SEQ ID NO:2;
- b) an isolated nucleic acid molecule comprising a nucleic acid sequence encoding a protein of (a), said nucleic acid sequence being operatively linked to a transcription control sequence;
- c) a product of drug discovery that has the biological activity of a protein comprising SEQ ID NO:2.
A method for the treatment of cancer, comprising contacting tumor cells in a patient with a compound, under conditions and for a time sufficient to permit the stimulation of the apoptotic process, said compound being selected from the group consisting of:
- a) an isolated protein comprising an amino acid sequence selected from the group consisting of:
- i) an amino acid sequence comprising SEQ ID NO:2;
- ii) a biologically active fragment of SEQ ID NO:2;
- and
- iii) an amino acid sequence that is at least about 50% identical to SEQ ID NO:2 and having a biological activity of SEQ ID NO:2;
- b) an isolated nucleic acid molecule comprising a nucleic acid sequence encoding a protein of (a), said nucleic acid sequence being operatively linked to a transcription control sequence;
- c) isolated antigen-presenting cells transfected with and expressing the isolated nucleic acid molecule of (b);
- and
- d) a product of drug discovery that has the biological activity of a protein comprising SEQ ID NO:2.
A method for reducing apoptosis in a cell, comprising contacting a cell intracellularly with a compound selected from the group consisting of:
- a) an antibody or antigen binding fragment thereof that selectively binds to and inhibits the activity of a protein comprising SEQ ID NO:2;
- b) an isolated protein that is at least about 50% identical and less than about 100% identical to SEQ ID NO:2, wherein said protein is an antagonist of a protein comprising SEQ ID NO:2;
- c) a fragment of SEQ ID NO:2 that has reduced apoptosis-inducing biological activity as compared to SEQ ID NO:2;
- d) an isolated nucleic acid sequence comprising at least 22 consecutive nucleotides of SEQ ID NO:1 or SEQ ID NO:5 and that hybridizes under highly stringent conditions to a gene encoding SEQ ID NO:2 and inhibits the expression of a protein comprising SEQ ID NO:2;
- and
- e) a product of drug discovery that inhibits the activity of a protein comprising SEQ ID NO:2 or the expression of a gene encoding a protein comprising SEQ ID NO:2.
A method to identify a regulator of apoptosis, comprising:
- a) contacting an apoptosis-inducing protein or nucleic acid molecule encoding said apoptosis-inducing protein with a putative regulatory compound, wherein said apoptosis-inducing protein comprises an amino acid sequence selected from the group consisting of:
- i) an amino acid sequence comprising SEQ ID NO:2;
- ii) a biologically active fragment of SEQ ID NO:2;
- and
- iii) an amino acid sequence that is at least about 50% identical to SEQ ID NO:2 and has a biological activity of SEQ ID NO:2;
- b) detecting whether said putative regulatory compound increases or decreases expression or activity of said protein or said nucleic acid molecule as compared to prior to contact with said compound;
- wherein compounds that increase or decrease the expression or activity of the protein or nucleic acid molecule, as compared to in the absence of said compound, indicates that said putative regulatory compound is a regulator of apoptosis.
Dependencies are inferred from the claim text.