KR910700258A - Mammalian GAP-43 Compositions Uses thereof - Google Patents

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Description

Mammalian GAP-43 Compositions Uses thereof

1 shows hybrid-selected transcription of GAP-43 cDNA. Figure 2 depicts the nucleotide sequence of GAP-43 and its intended amino acid sequence. 3 shows the regulation of GAP-43 expression in PC12 cells. Pass PC12 cells in RPMI medium containing 10% horse serum and 5% bovine serum.

Claims (77)

Substantially pure mammalian GAP-43 or a functional derivative thereof. The composition of claim 1, wherein the mammalian GAP-43 is rat GAP-43. The composition of claim 1, wherein the mammalian GAP-43 is human GAP-43. The composition of claim 2, wherein rat GAP-43 has an amino acid sequence substantially similar to that shown in FIG. 2. 4. The composition of claim 3, wherein human GAP-43 has an amino acid sequence substantially similar to that shown in Figure 5A. A polypeptide, or functional derivative thereof, containing an amino acid sequence substantially similar to that shown in FIG. A polypeptide, or functional derivative thereof, containing an amino acid sequence substantially similar to that shown in FIG. 5A. CDNA containing a nucleotide substantially similar to that shown in FIG. 2, or a functional derivative thereof. CDNA, or functional derivative thereof, containing a nucleotide substantially similar to that shown in FIG. 5A. A DNA expression vector containing the cDNA of claim 8. A DNA expression vector containing the cDNA of claim 9. A host cell transfected with the vector of claim 10. The host cell of claim 12, wherein the cell is selected from the group consisting of prokaryotic and eukaryotic cells. GAP-43 produced by the cell of claim 13, or a functional derivative thereof. Prokaryotic or eukaryotic cells are transfected with a vector containing GD-43 of a cDNA encoding-mammal, and the host cells are cultured in an appropriate medium under conditions such that the mammalian GAP-43 is expressed, and the mammalian GAP from the medium -43 is isolated, thereby producing a mammalian GAP-43 or a functional derivative thereof. An antibody directed against GAP-43 or a functional or chemical derivative thereof of a mammal. Antibodies directed against GAP-43 or a functional or chemical derivative thereof of the rat. Antibodies directed against human GAP-43 or functional or chemical derivatives thereof. The antibody of claim 16, 17 or 18, wherein the antibody is selected from the group consisting of monoclonal antibodies and polyclonal antibodies. The antibody of claim 19, wherein the antibody is detectably labeled. The antibody of claim 19, wherein the antibody is therapeutically labeled. A pharmaceutical composition comprising the composition of any one of claims 1-7 or 14 together with a pharmaceutically acceptable carrier. The composition of claim 22, further comprising one or more therapeutically effective agents. A sample suspected of containing GAP-43 was contacted with the antibody of claim 20, the sample was incubated with the antibody to form a GAP-43 antibody complex, and then the antibody which formed the complex was isolated from the antibody which did not form the complex. And measuring or detecting the labeled complex forming antibody. A kit for measuring or detecting GAP-43, comprising a carrier partitioned to accommodate at least one container containing a detectably labeled antibody against GAP-43 in a closed compartment. The kit of claim 25, wherein the antibody is selected from the group consisting of polyclonal and monoclonal. Exposing the cell to an effective amount of nerve growth factor, thereby inducing expression of GAP-43 in the cell. The method of claim 27, wherein the cells are neurons. The method of claim 28, wherein the cell is directly exposed to nerve growth factor. A method of enhancing expression of GAP-43 in a cell, characterized by introducing a DNA expression vector containing cDNA encoding GAP-43 into a cell. The method of claim 30, wherein the vector is introduced into the cell by transfection, transduction or direct microinjection. 32. A method for promoting structural remodeling of neurons, characterized by inducing GAP-43 expression in cells by the method of any one of claims 28-31. Inducing expression of GAP-43 in and around the damaged nerve tissue, thereby promoting recovery of the damaged nerve tissue. 34. The method of claim 33, wherein the nerve injury is caused by an incomplete fracture with ischemia, transient ischemia without an incomplete fracture, hypoxia, oxygen deficiency, oxygen deficiency encephalitis, hyporeflux, or seizure. A method for regulating structural reconstitution of neuronal cells, characterized by exposing the cells to an effective amount of at least one substance selected from the group consisting of nerve growth factors, steroids and functional derivatives thereof. A method for regulating synaptic formability of neuronal cells, characterized by exposing the cells to an effective amount of at least one substance selected from the group consisting of nerve growth factors, steroids and functional derivatives thereof. A method for controlling microenvironment of neuronal cells, characterized by exposing an effective amount of cells of at least one substance selected from the group consisting of nerve growth factors, steroids and functional derivatives thereof. Exposing the cells to at least one steroid effective amount, thereby inhibiting the expression of GAP-43 in mammalian neurons. Exposing the cells to at least one steroid effective amount, thereby inhibiting the expression of GAP-43 in fully differentiated mammalian neuronal cells. 40. The method of any one of claims 35-39, wherein the steroid is a cortical steroid. 41. The method of claim 40, wherein the cortical steroid is selected from the group consisting of mineral corticoids and glucocolticoids. 42. The method of claim 41, wherein the mineral corticoid is selected from the group consisting of dexamethasone, corticosterone, aldosterone, and progesterone. Exposing the cells to an effective amount of cyclohexamide, thereby enhancing steroidal inhibition of GAP-43 expression of mammalian neuronal cells exposed to steroids. 32. The method of claim 30 or 31, wherein the cells are non-neuronal cells. Nucleotide sequence aatg ctg tgc tgt atg aga acc aaa cag or cDNA encoding a membrane-target membrane peptide containing a functional derivative thereof. I. MET LEU CYS CYS MET MARG ARG THR LYS GLN; II. MET LEU CYS CYS MET ARG ARG THR LYS; III. MET LEU CYS CYS MET ARG ARG THR; IV. MET LEU CYS CYS MET ARG ARG; V. MET LEU CYS CYS MET ARG; VI. MET LEU CYS CYS MET; VII. MET LEU CYS CYS; And VIII. Membrane-target membrane peptide containing an amino acid sequence selected from the group consisting of functional derivatives thereof. I. MET LEU CYS CYS MET MARG ARG THR LYS GLN; II. MET LEU CYS CYS MET ARG ARG THR LYS; III. MET LEU CYS CYS MET ARG ARG THR; IV. MET LEU CYS CYS MET ARG ARG; V. MET LEU CYS CYS MET ARG; VI. MET LEU CYS CYS MET; VII. MET LEU CYS CYS; And VIII. DNA encoding a membrane-targeting peptide containing a nucleotide encoding an amino acid sequence selected from the group consisting of other functional derivatives. A structural gene or fragment thereof containing a nucleotide encoding a membrane-targeting peptide having an amino-terminal sequence of claim 46. A protein or peptide containing a membrane-targeting peptide having the sequence of claim 46 at the amino terminus. (a) linking the membrane-targeting peptide containing the amino acid sequence of claim 46 to the amino terminus of the protein or peptide, and (b) introducing the resulting protein or peptide containing the membrane-targeting domain into the cell, (Wherein the protein or peptide of step (b) functions directly on the cell's membrane by means of the membrane-targeting domain), a method of directly acting the protein or peptide of interest on the cell membrane. 51. The method of claim 50, wherein the cells are selected from the group consisting of neuronal- or nonneuronal-cells. The method of claim 51, wherein the resulting protein and peptide of step (b) in neuronal cells act directly on the growth dendritic region of the cell. Has substantially the specificity of MAb anti-GAP-43 (H5) or a functional or chemical derivative thereof, wherein the MAb anti-GAP-43 (H5) is produced in hybridoma strain H5, wherein the hybridoma strain is ATCC _____ Monoclonal antibodies. Hybridoma strain H5, ATCC ____, functional or chemical derivatives thereof. Nucleotide sequence of FIG. 13 encoding genomic GAP-43, or a functional or chemical derivative thereof. Nucleotide sequence encoding GAP-43, or a functional or chemical derivative thereof. Long homopurine-homopyrimidine extension with multiple starting and condensate pit-1 linkages, without TATA-box and Consus Sp-1 linkages, to create triple-chain (H-DNA) structures A promoter substantially identical to that shown in FIG. 14, characterized by a call. A structural gene or fragment thereof which, at its amino terminus, contains the nucleotide sequence of claim 56 or 57 or a functional or chemical derivative thereof. A DNA expression vector containing the structural gene of claim 58. A host cell transfected with the vector of claim 59. Internal regulatory protein (IRP). XXIII. IRP peptide containing an amino acid sequence selected from the group consisting of functional derivatives thereof. The nucleotide sequence encoding the IRP peptide of claim 62. IRP peptide with consensus amino acid sequence hydrophobic-1eu-cys-cys-x-basic-basic or a functional derivative thereof. 65. The IRP peptide of claim 62 or 64 wherein the cysteine is palmitic acid treated. A method for stimulating the binding activity of a protein of interest, characterized by introducing an effective amount of the IRP peptide into an environment containing the protein of interest and its binding substrate. 67. The method of claim 66, wherein the protein of interest is a G protein. The method of claim 67, wherein the G protein is G ₈ . 67. The method of claim 66, wherein the IRP peptide is the peptide of claim 62, 64 or 65. 67. The method of claim 66, wherein the environment is inside viable cells. The method of claim 70, wherein the cells are central or peripheral neurons. The method of claim 67, wherein the binding substrate is GTP. A monoclonal antibody, or a functional or chemical derivative thereof, directed against the IRP of claim 62 or 63 and optionally labeled for detection or treatment. A pharmaceutical composition comprising the IRP peptide of claim 62 or 74 together with a pharmaceutically acceptable carrier and optionally containing one or more therapeutic benefit agents. A pharmaceutical composition comprising the monoclonal antibody of claim 73 together with a pharmaceutically acceptable carrier and optionally containing one or more therapeutic benefit agents. 76. A method of modulating structural reconstitution of a neuron, characterized by administering an effective amount of the composition of claim 74 or 75 to the cell. Neuronal growth-characterized by binding specifically to GAP-43, molecular weight 39,000, eluting in a single band when using EDTA in 50 mM Tris buffer, and not reacting with polyclonal antibodies against G ₈ Related proteins. ※ Note: The disclosure is based on the initial application.