Caspase-mediated Parkin Cleavage in Apoptotic Cell Death

The parkin protein is important for the survival of the neurons that degenerate in Parkinson's disease as demonstrated by disease-causing lesions in the parkin gene. The Chinese hamster ovary and the SH-SY5Y cell line stably expressing recombinant human parkin combined with epitope-specific parkin antibodies were used to investigate the proteolytic processing of human parkin during apoptosis by immunoblotting. Parkin is cleaved during apoptosis induced by okadaic acid, staurosporine, and camptothecin, thereby generating a 38-kDa C-terminal fragment and a 12-kDa N-terminal fragment. The cleavage was not significantly affected by the disease-causing mutations K161N, G328E, T415N, and G430D and the polymorphism R366W. Parkin and its 38-kDa proteolytic fragment is preferentially associated with vesicles, thereby indicating that cleavage is a membrane-associated event. The proteolysis is sensitive to inhibitors of caspases. The cleavage site was mapped by site-directed mutagenesis of potential aspartic residues and revealed that mutation of Asp-126 alone abrogated the parkin cleavage. The tetrapeptide aldehyde LHTD-CHO, representing the amino acid sequence N-terminal to the putative cleavage site was an efficient inhibitor of parkin cleavage. This suggests that parkin function is compromised in neuropathological states associated with an increased caspase activation, thereby further adding to the cellular stress. The parkin protein is important for the survival of the neurons that degenerate in Parkinson's disease as demonstrated by disease-causing lesions in the parkin gene. The Chinese hamster ovary and the SH-SY5Y cell line stably expressing recombinant human parkin combined with epitope-specific parkin antibodies were used to investigate the proteolytic processing of human parkin during apoptosis by immunoblotting. Parkin is cleaved during apoptosis induced by okadaic acid, staurosporine, and camptothecin, thereby generating a 38-kDa C-terminal fragment and a 12-kDa N-terminal fragment. The cleavage was not significantly affected by the disease-causing mutations K161N, G328E, T415N, and G430D and the polymorphism R366W. Parkin and its 38-kDa proteolytic fragment is preferentially associated with vesicles, thereby indicating that cleavage is a membrane-associated event. The proteolysis is sensitive to inhibitors of caspases. The cleavage site was mapped by site-directed mutagenesis of potential aspartic residues and revealed that mutation of Asp-126 alone abrogated the parkin cleavage. The tetrapeptide aldehyde LHTD-CHO, representing the amino acid sequence N-terminal to the putative cleavage site was an efficient inhibitor of parkin cleavage. This suggests that parkin function is compromised in neuropathological states associated with an increased caspase activation, thereby further adding to the cellular stress. ubiquitin ligase ubiquitin-activating enzyme ubiquitin-conjugating enzyme Chinese hamster ovary okadaic acid Parkinson's disease is the second most common neurodegenerative disorder, and its symptoms arise primarily from a rather selective loss of dopaminergic neurons in the substantia nigra of the brain stem (1Lang A.E. Lozano A.M. N. Engl. J. Med. 1998; 339: 1044-1053Crossref PubMed Scopus (1749) Google Scholar). Lesions in the parkin gene on chromosome 6 are responsible for a large number of patients affected by early onset Parkinson's disease (2Kitada T. Asakawa S. Hattori N. Matsumine H. Yamamura Y. Minoshima S. Yokochi M. Mizuno Y. Shimizu N. Nature. 1998; 392: 605-608Crossref PubMed Scopus (4123) Google Scholar, 3Lücking C.B. Durr A. Bonifati V. De Vaughan J. Michele G. Gasser T. Harhangi B.S. Meco G. Denefle P. Wood N.W. Agid Y. Brice A. N. Engl. J. Med. 2000; 342: 1560-1567Crossref PubMed Scopus (1237) Google Scholar). The parkin gene encodes the intracellular parkin protein that consists of 465 amino acids with an N-terminal ubiquitin-like domain and a C-terminal domain with two Ring finger motifs (2Kitada T. Asakawa S. Hattori N. Matsumine H. Yamamura Y. Minoshima S. Yokochi M. Mizuno Y. Shimizu N. Nature. 1998; 392: 605-608Crossref PubMed Scopus (4123) Google Scholar). Ring finger domains are present in one class of E31 ubiquitin ligases (4Kornitzer D. Ciechanover A. J. Cell. Physiol. 2000; 182: 1-11Crossref PubMed Scopus (231) Google Scholar), and parkin exhibits ubiquitin ligase activity although it is unclear whether it is a bona fide E3 ubiquitin ligase or exists as part of a multisubunit ubiquitin ligase complex (5Shimura H. Hattori N. Kubo S. Mizuno Y. Asakawa S. Minoshima S. Shimizu N. Iwai K. Chiba T. Tanaka K. Suzuki T. Nat. Genet. 2000; 25: 302-305Crossref PubMed Scopus (1681) Google Scholar, 6Imai Y. Soda M. Takahashi R. J. Biol. Chem. 2000; 275: 35661-35664Abstract Full Text Full Text PDF PubMed Scopus (650) Google Scholar, 7Zhang Y Gao J. Chung K.K.K. Huang H. Dawson V. Dawson T.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: PubMed Scopus Google Scholar). The C-terminal Ring finger domain the to ubiquitin (2Kitada T. Asakawa S. Hattori N. Matsumine H. Yamamura Y. Minoshima S. Yokochi M. Mizuno Y. Shimizu N. Nature. 1998; 392: 605-608Crossref PubMed Scopus (4123) Google Y Gao J. Chung K.K.K. Huang H. Dawson V. Dawson T.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: PubMed Scopus Google is responsible for the of intracellular protein and on the of two enzyme the ubiquitin and the Ciechanover A. 1998; PubMed Scopus Google Scholar). of is by by the ubiquitin-activating enzyme ubiquitin and ubiquitin ligases The E3 ubiquitin ligases function as that the in the of the or by the ubiquitin to the The from to cellular of the cell thereby by the The of the parkin gene is during protein as part of the protein and parkin protein Y. Soda M. Takahashi R. J. Biol. Chem. 2000; 275: 35661-35664Abstract Full Text Full Text PDF PubMed Scopus (650) Google Scholar, Y. Soda M. H. Hattori N. Mizuno Y. Takahashi R. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). disease-causing lesions in the parkin gene in a loss of ubiquitin ligase function (5Shimura H. Hattori N. Kubo S. Mizuno Y. Asakawa S. Minoshima S. Shimizu N. Iwai K. Chiba T. Tanaka K. Suzuki T. Nat. Genet. 2000; 25: 302-305Crossref PubMed Scopus (1681) Google Scholar, 7Zhang Y Gao J. Chung K.K.K. Huang H. Dawson V. Dawson T.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: PubMed Scopus Google Scholar, Y. Soda M. H. Hattori N. Mizuno Y. Takahashi R. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). The loss of function a of the dopaminergic neurons in the substantia nigra parkin in the H. Hattori N. Kubo S. M. T. Matsumine H. Asakawa S. Minoshima S. Yamamura Y. Shimizu N. Mizuno Y. PubMed Scopus Google Scholar, J. G. PubMed Scopus Google Scholar, N. A. Agid Y. R. Brice A. J. 2000; PubMed Scopus Google Scholar, M. V. A. A. A. J. S. R. Agid Y. Brice A. J. PubMed Scopus Google Scholar). This that parkin a function for the survival of of that on parkin function potential to the in Parkinson's a of that the of apoptosis and an for aspartic residues in the of J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, 2000; PubMed Scopus Google Scholar, M. M. J. 2000; PubMed Scopus Google Scholar). cell is a caspase is by cellular and Y. 1998; PubMed Scopus Google Scholar). caspase is with cellular survival S. Nature. PubMed Scopus Google Scholar, R. Nature. PubMed Scopus Google Scholar), and a caspase activity that caspase to a in neurodegenerative Y. A. J. 2000; PubMed Google Scholar). This is by the of or proteolytic in of brain affected by Parkinson's disease A. S. S. A. H. A. M. Agid Y. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: PubMed Scopus Google and disease A. H. J. Full Text Full Text PDF PubMed Scopus Google Scholar, J. Full Text Full Text PDF PubMed Scopus Google Scholar). by in the of common neurodegenerative the of parkin and that parkin is cleaved during apoptosis by Asp-126 the cellular parkin cleavage site in Chinese hamster ovary and SH-SY5Y dopaminergic cell as by site-directed of parkin Asp-126 the disease-causing of amino acid residues C.B. Durr A. Bonifati V. De Vaughan J. Michele G. Gasser T. Harhangi B.S. Meco G. Denefle P. Wood N.W. Agid Y. Brice A. N. Engl. J. Med. 2000; 342: 1560-1567Crossref PubMed Scopus (1237) Google and is with a parkin proteolysis parkin and a 38-kDa C-terminal fragment as demonstrated by its of antibodies recombinant human parkin and a to the C-terminal amino acid residues of human parkin and the abrogated of the 38-kDa fragment in the of inhibitors of caspases. Parkin cleavage is induced by as of apoptosis as the protein inhibitor the inhibitor staurosporine, and the inhibitor This that parkin cleavage is associated to apoptosis in and not the of a cellular of caspase activity in the cleavage is by of two caspase inhibitors the thereby a caspase as of or as responsible for the cleavage. the of cleavage by site-directed mutagenesis of Asp-126 to and that the an in the the for an in the is by the efficient of the cleavage by the tetrapeptide aldehyde on the amino acid sequence N-terminal to the putative in parkin caspase proteolysis of was as a the as a cleavage site in parkin Y. of the for 2000; Scholar), site in cellular on site-directed The of the parkin caspase from the of the inhibitors as a cell The sequence N-terminal to the cleavage site not to of the for caspase cleavage cleaved with by and as on the by T. M. S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). The of the responsible caspase to the of a cell caspase or the of cell from caspase gene of that parkin is a protein as demonstrated T. S. H. Y. Asakawa S. Minoshima S. Shimizu N. Mizuno Y. Hattori N. J. PubMed Scopus Google Scholar). This is in the the 38-kDa fragment This that the parkin cleavage or the A. A.M. 2000; PubMed Scopus Google T. H. N. J. J. Nature. 2000; PubMed Scopus Google of parkin Asp-126 the ubiquitin-like domain from the large the Ring domain This is with a parkin enzyme on that a mutation in the ubiquitin-like domain or of and in the loss of amino acid residues early onset on parkin C.B. Durr A. Bonifati V. De Vaughan J. Michele G. Gasser T. Harhangi B.S. Meco G. Denefle P. Wood N.W. Agid Y. Brice A. N. Engl. J. Med. 2000; 342: 1560-1567Crossref PubMed Scopus (1237) Google Scholar, N. C.B. S. Durr A. De Bonifati V. Michele G. S. Vaughan Gasser T. R. Harhangi B.S. Wood N.W. A. Meco G. Denefle P. Agid Y. Brice A. Genet. PubMed Scopus Google Scholar). and cellular that of the ubiquitin-like domain in ubiquitin ligase activity and of parkin (5Shimura H. Hattori N. Kubo S. Mizuno Y. Asakawa S. Minoshima S. Shimizu N. Iwai K. Chiba T. Tanaka K. Suzuki T. Nat. Genet. 2000; 25: 302-305Crossref PubMed Scopus (1681) Google Scholar, 6Imai Y. Soda M. Takahashi R. J. Biol. Chem. 2000; 275: 35661-35664Abstract Full Text Full Text PDF PubMed Scopus (650) Google Scholar, 7Zhang Y Gao J. Chung K.K.K. Huang H. Dawson V. Dawson T.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: PubMed Scopus Google Scholar, Y. Soda M. H. Hattori N. Mizuno Y. Takahashi R. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar, H. Hattori N. A. R. Mizuno Y. PubMed Scopus Google Scholar). of the expressing the N-terminal acid parkin its in the This suggests that it function in by the Y. of the for 2000; for associated to the in cell and that and proteolytic are present in neurons in brain from M. A. PubMed Scopus Google Scholar, T. P. 2000; PubMed Scopus Google and affected by disease Y. 1998; PubMed Scopus Google Scholar, 2000; PubMed Scopus Google and neurodegenerative S. Nature. PubMed Scopus Google Scholar, R. Nature. PubMed Scopus Google Scholar, H. Hattori N. A. R. Mizuno Y. PubMed Scopus Google Scholar, M. A. PubMed Scopus Google Scholar, D. Y. Huang J. A. D. M. H. S. Cell. 97: Full Text Full Text PDF PubMed Scopus Google Scholar, M. J. PubMed Scopus Google are to for The neurons are that caspase activity is with survival of compromised Y. 1998; PubMed Scopus Google Scholar, S. Nature. PubMed Scopus Google Scholar, R. Nature. PubMed Scopus Google Scholar, M. J. PubMed Scopus Google to a 2000; PubMed Scopus Google Scholar). This is by cell neurons of as with cell that M. M. J. 2000; PubMed Scopus Google Scholar). H. Hattori N. Kubo S. M. T. Matsumine H. Asakawa S. Minoshima S. Yamamura Y. Shimizu N. Mizuno Y. PubMed Scopus Google the of an of in brain from patients with is in substantia with antibodies are to a parkin cleavage with a proteolysis in of substantia nigra not The of putative in cleavage of parkin the of antibodies that parkin as protein D. Y. Huang J. A. D. M. H. S. Cell. 97: Full Text Full Text PDF PubMed Scopus Google cleavage to a in neurons caspase activation, by in Parkinson's parkin function that the cellular and to further caspase the neurons a is a demonstrated in a of of the of the protein was by of the symptoms of the A. R. Cell. 2000; Full Text Full Text PDF PubMed Scopus Google not of the mutation as with parkin the with staurosporine, and not the of an as the of parkin demonstrated to rather parkin cleavage a in Parkinson's disease the of dopaminergic neurons that the of of used in Parkinson's are to a as by the large cell loss M. P. H. P. Nat. Med. PubMed Scopus Google Scholar). of parkin Asp-126 in an for the survival the mutation not the activity of Parkinson's disease is the second most common neurodegenerative disorder, and its symptoms arise primarily from a rather selective loss of dopaminergic neurons in the substantia nigra of the brain stem (1Lang A.E. Lozano A.M. N. Engl. J. Med. 1998; 339: 1044-1053Crossref PubMed Scopus (1749) Google Scholar). Lesions in the parkin gene on chromosome 6 are responsible for a large number of patients affected by early onset Parkinson's disease (2Kitada T. Asakawa S. Hattori N. Matsumine H. Yamamura Y. Minoshima S. Yokochi M. Mizuno Y. Shimizu N. Nature. 1998; 392: 605-608Crossref PubMed Scopus (4123) Google Scholar, 3Lücking C.B. Durr A. Bonifati V. De Vaughan J. Michele G. Gasser T. Harhangi B.S. Meco G. Denefle P. Wood N.W. Agid Y. Brice A. N. Engl. J. Med. 2000; 342: 1560-1567Crossref PubMed Scopus (1237) Google Scholar). The parkin gene encodes the intracellular parkin protein that consists of 465 amino acids with an N-terminal ubiquitin-like domain and a C-terminal domain with two Ring finger motifs (2Kitada T. Asakawa S. Hattori N. Matsumine H. Yamamura Y. Minoshima S. Yokochi M. Mizuno Y. Shimizu N. Nature. 1998; 392: 605-608Crossref PubMed Scopus (4123) Google Scholar). Ring finger domains are present in one class of E31 ubiquitin ligases (4Kornitzer D. Ciechanover A. J. Cell. Physiol. 2000; 182: 1-11Crossref PubMed Scopus (231) Google Scholar), and parkin exhibits ubiquitin ligase activity although it is unclear whether it is a bona fide E3 ubiquitin ligase or exists as part of a multisubunit ubiquitin ligase complex (5Shimura H. Hattori N. Kubo S. Mizuno Y. Asakawa S. Minoshima S. Shimizu N. Iwai K. Chiba T. Tanaka K. Suzuki T. Nat. Genet. 2000; 25: 302-305Crossref PubMed Scopus (1681) Google Scholar, 6Imai Y. Soda M. Takahashi R. J. Biol. Chem. 2000; 275: 35661-35664Abstract Full Text Full Text PDF PubMed Scopus (650) Google Scholar, 7Zhang Y Gao J. Chung K.K.K. Huang H. Dawson V. Dawson T.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: PubMed Scopus Google Scholar). The C-terminal Ring finger domain the to ubiquitin (2Kitada T. Asakawa S. Hattori N. Matsumine H. Yamamura Y. Minoshima S. Yokochi M. Mizuno Y. Shimizu N. Nature. 1998; 392: 605-608Crossref PubMed Scopus (4123) Google Y Gao J. Chung K.K.K. Huang H. Dawson V. Dawson T.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: PubMed Scopus Google Scholar). The is responsible for the of intracellular protein and on the of two enzyme the ubiquitin and the Ciechanover A. 1998; PubMed Scopus Google Scholar). of is by by the ubiquitin-activating enzyme ubiquitin and ubiquitin ligases The E3 ubiquitin ligases function as that the in the of the or by the ubiquitin to the The from to cellular of the cell thereby by the The of the parkin gene is during protein as part of the protein and parkin protein Y. Soda M. Takahashi R. J. Biol. Chem. 2000; 275: 35661-35664Abstract Full Text Full Text PDF PubMed Scopus (650) Google Scholar, Y. Soda M. H. Hattori N. Mizuno Y. Takahashi R. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). disease-causing lesions in the parkin gene in a loss of ubiquitin ligase function (5Shimura H. Hattori N. Kubo S. Mizuno Y. Asakawa S. Minoshima S. Shimizu N. Iwai K. Chiba T. Tanaka K. Suzuki T. Nat. Genet. 2000; 25: 302-305Crossref PubMed Scopus (1681) Google Scholar, 7Zhang Y Gao J. Chung K.K.K. Huang H. Dawson V. Dawson T.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: PubMed Scopus Google Scholar, Y. Soda M. H. Hattori N. Mizuno Y. Takahashi R. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). The loss of function a of the dopaminergic neurons in the substantia nigra parkin in the H. Hattori N. Kubo S. M. T. Matsumine H. Asakawa S. Minoshima S. Yamamura Y. Shimizu N. Mizuno Y. PubMed Scopus Google Scholar, J. G. PubMed Scopus Google Scholar, N. A. Agid Y. R. Brice A. J. 2000; PubMed Scopus Google Scholar, M. V. A. A. A. J. S. R. Agid Y. Brice A. J. PubMed Scopus Google Scholar). This that parkin a function for the survival of of that on parkin function potential to the in Parkinson's a of that the of apoptosis and an for aspartic residues in the of J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, 2000; PubMed Scopus Google Scholar, M. M. J. 2000; PubMed Scopus Google Scholar). cell is a caspase is by cellular and Y. 1998; PubMed Scopus Google Scholar). caspase is with cellular survival S. Nature. PubMed Scopus Google Scholar, R. Nature. PubMed Scopus Google Scholar), and a caspase activity that caspase to a in neurodegenerative Y. A. J. 2000; PubMed Google Scholar). This is by the of or proteolytic in of brain affected by Parkinson's disease A. S. S. A. H. A. M. Agid Y. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: PubMed Scopus Google and disease A. H. J. Full Text Full Text PDF PubMed Scopus Google Scholar, J. Full Text Full Text PDF PubMed Scopus Google Scholar). by in the of common neurodegenerative the of parkin and that parkin is cleaved during apoptosis by Asp-126 the cellular parkin cleavage site in Chinese hamster ovary and SH-SY5Y dopaminergic cell as by site-directed of parkin Asp-126 the disease-causing of amino acid residues C.B. Durr A. Bonifati V. De Vaughan J. Michele G. Gasser T. Harhangi B.S. Meco G. Denefle P. Wood N.W. Agid Y. Brice A. N. Engl. J. Med. 2000; 342: 1560-1567Crossref PubMed Scopus (1237) Google and is with a parkin proteolysis parkin and a 38-kDa C-terminal fragment as demonstrated by its of antibodies recombinant human parkin and a to the C-terminal amino acid residues of human parkin and the abrogated of the 38-kDa fragment in the of inhibitors of caspases. Parkin cleavage is induced by as of apoptosis as the protein inhibitor the inhibitor staurosporine, and the inhibitor This that parkin cleavage is associated to apoptosis in and not the of a cellular of caspase activity in the cleavage is by of two caspase inhibitors the thereby a caspase as of or as responsible for the cleavage. the of cleavage by site-directed mutagenesis of Asp-126 to and that the an in the the for an in the is by the efficient of the cleavage by the tetrapeptide aldehyde on the amino acid sequence N-terminal to the putative in parkin caspase proteolysis of was as a the as a cleavage site in parkin Y. of the for 2000; Scholar), site in cellular on site-directed The of the parkin caspase from the of the inhibitors as a cell The sequence N-terminal to the cleavage site not to of the for caspase cleavage cleaved with by and as on the by T. M. S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). The of the responsible caspase to the of a cell caspase or the of cell from caspase gene of that parkin is a protein as demonstrated T. S. H. Y. Asakawa S. Minoshima S. Shimizu N. Mizuno Y. Hattori N. J. PubMed Scopus Google Scholar). This is in the the 38-kDa fragment This that the parkin cleavage or the A. A.M. 2000; PubMed Scopus Google T. H. N. J. J. Nature. 2000; PubMed Scopus Google of parkin Asp-126 the ubiquitin-like domain from the large the Ring domain This is with a parkin enzyme on that a mutation in the ubiquitin-like domain or of and in the loss of amino acid residues early onset on parkin C.B. Durr A. Bonifati V. De Vaughan J. Michele G. Gasser T. Harhangi B.S. Meco G. Denefle P. Wood N.W. Agid Y. Brice A. N. Engl. J. Med. 2000; 342: 1560-1567Crossref PubMed Scopus (1237) Google Scholar, N. C.B. S. Durr A. De Bonifati V. Michele G. S. Vaughan Gasser T. R. Harhangi B.S. Wood N.W. A. Meco G. Denefle P. Agid Y. Brice A. Genet. PubMed Scopus Google Scholar). and cellular that of the ubiquitin-like domain in ubiquitin ligase activity and of parkin (5Shimura H. Hattori N. Kubo S. Mizuno Y. Asakawa S. Minoshima S. Shimizu N. Iwai K. Chiba T. Tanaka K. Suzuki T. Nat. Genet. 2000; 25: 302-305Crossref PubMed Scopus (1681) Google Scholar, 6Imai Y. Soda M. Takahashi R. J. Biol. Chem. 2000; 275: 35661-35664Abstract Full Text Full Text PDF PubMed Scopus (650) Google Scholar, 7Zhang Y Gao J. Chung K.K.K. Huang H. Dawson V. Dawson T.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: PubMed Scopus Google Scholar, Y. Soda M. H. Hattori N. Mizuno Y. Takahashi R. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar, H. Hattori N. A. R. Mizuno Y. PubMed Scopus Google Scholar). of the expressing the N-terminal acid parkin its in the This suggests that it function in by the Y. of the for 2000; for associated to the in cell and that and proteolytic are present in neurons in brain from M. A. PubMed Scopus Google Scholar, T. P. 2000; PubMed Scopus Google and affected by disease Y. 1998; PubMed Scopus Google Scholar, 2000; PubMed Scopus Google and neurodegenerative S. Nature. PubMed Scopus Google Scholar, R. Nature. PubMed Scopus Google Scholar, H. Hattori N. A. R. Mizuno Y. PubMed Scopus Google Scholar, M. A. PubMed Scopus Google Scholar, D. Y. Huang J. A. D. M. H. S. Cell. 97: Full Text Full Text PDF PubMed Scopus Google Scholar, M. J. PubMed Scopus Google are to for The neurons are that caspase activity is with survival of compromised Y. 1998; PubMed Scopus Google Scholar, S. Nature. PubMed Scopus Google Scholar, R. Nature. PubMed Scopus Google Scholar, M. J. PubMed Scopus Google to a 2000; PubMed Scopus Google Scholar). This is by cell neurons of as with cell that M. M. J. 2000; PubMed Scopus Google Scholar). H. Hattori N. Kubo S. M. T. Matsumine H. Asakawa S. Minoshima S. Yamamura Y. Shimizu N. Mizuno Y. PubMed Scopus Google the of an of in brain from patients with is in substantia with antibodies are to a parkin cleavage with a proteolysis in of substantia nigra not The of putative in cleavage of parkin the of antibodies that parkin as protein D. Y. Huang J. A. D. M. H. S. Cell. 97: Full Text Full Text PDF PubMed Scopus Google cleavage to a in neurons caspase activation, by in Parkinson's parkin function that the cellular and to further caspase the neurons a is a demonstrated in a of of the of the protein was by of the symptoms of the A. R. Cell. 2000; Full Text Full Text PDF PubMed Scopus Google not of the mutation as with parkin the with staurosporine, and not the of an as the of parkin demonstrated to rather parkin cleavage a in Parkinson's disease the of dopaminergic neurons that the of of used in Parkinson's are to a as by the large cell loss M. P. H. P. Nat. Med. PubMed Scopus Google Scholar). of parkin Asp-126 in an for the survival the mutation not the activity of proteolysis parkin and a 38-kDa C-terminal fragment as demonstrated by its of antibodies recombinant human parkin and a to the C-terminal amino acid residues of human parkin and the abrogated of the 38-kDa fragment in the of inhibitors of caspases. Parkin cleavage is induced by as of apoptosis as the protein inhibitor the inhibitor staurosporine, and the inhibitor This that parkin cleavage is associated to apoptosis in and not the of a cellular of caspase activity in the cleavage is by of two caspase inhibitors the thereby a caspase as of or as responsible for the cleavage. the of cleavage by site-directed mutagenesis of Asp-126 to and that the an in the the for an in the is by the efficient of the cleavage by the tetrapeptide aldehyde on the amino acid sequence N-terminal to the putative in parkin caspase proteolysis of was as a the as a cleavage site in parkin Y. of the for 2000; Scholar), site in cellular on site-directed The of the parkin caspase from the of the inhibitors as a cell The sequence N-terminal to the cleavage site not to of the for caspase cleavage cleaved with by and as on the by T. M. S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). The of the responsible caspase to the of a cell caspase or the of cell from caspase gene of that parkin is a protein as demonstrated T. S. H. Y. Asakawa S. Minoshima S. Shimizu N. Mizuno Y. Hattori N. J. PubMed Scopus Google Scholar). This is in the the 38-kDa fragment This that the parkin cleavage or the A. A.M. 2000; PubMed Scopus Google T. H. N. J. J. Nature. 2000; PubMed Scopus Google Scholar). of parkin Asp-126 the ubiquitin-like domain from the large the Ring domain This is with a parkin enzyme on that a mutation in the ubiquitin-like domain or of and in the loss of amino acid residues early onset on parkin C.B. Durr A. Bonifati V. De Vaughan J. Michele G. Gasser T. Harhangi B.S. Meco G. Denefle P. Wood N.W. Agid Y. Brice A. N. Engl. J. Med. 2000; 342: 1560-1567Crossref PubMed Scopus (1237) Google Scholar, N. C.B. S. Durr A. De Bonifati V. Michele G. S. Vaughan Gasser T. R. Harhangi B.S. Wood N.W. A. Meco G. Denefle P. Agid Y. Brice A. Genet. PubMed Scopus Google Scholar). and cellular that of the ubiquitin-like domain in ubiquitin ligase activity and of parkin (5Shimura H. Hattori N. Kubo S. Mizuno Y. Asakawa S. Minoshima S. Shimizu N. Iwai K. Chiba T. Tanaka K. Suzuki T. Nat. Genet. 2000; 25: 302-305Crossref PubMed Scopus (1681) Google Scholar, 6Imai Y. Soda M. Takahashi R. J. Biol. Chem. 2000; 275: 35661-35664Abstract Full Text Full Text PDF PubMed Scopus (650) Google Scholar, 7Zhang Y Gao J. Chung K.K.K. Huang H. Dawson V. Dawson T.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: PubMed Scopus Google Scholar, Y. Soda M. H. Hattori N. Mizuno Y. Takahashi R. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar, H. Hattori N. A. R. Mizuno Y. PubMed Scopus Google Scholar). of the expressing the N-terminal acid parkin its in the This suggests that it function in by the Y. of the for 2000; Scholar). for associated to the in cell and that and proteolytic are present in neurons in brain from M. A. PubMed Scopus Google Scholar, T. P. 2000; PubMed Scopus Google and affected by disease Y. 1998; PubMed Scopus Google Scholar, 2000; PubMed Scopus Google and neurodegenerative S. Nature. PubMed Scopus Google Scholar, R. Nature. PubMed Scopus Google Scholar, H. Hattori N. A. R. Mizuno Y. PubMed Scopus Google Scholar, M. A. PubMed Scopus Google Scholar, D. Y. Huang J. A. D. M. H. S. Cell. 97: Full Text Full Text PDF PubMed Scopus Google Scholar, M. J. PubMed Scopus Google are to for The neurons are that caspase activity is with survival of compromised Y. 1998; PubMed Scopus Google Scholar, S. Nature. PubMed Scopus Google Scholar, R. Nature. PubMed Scopus Google Scholar, M. J. PubMed Scopus Google to a 2000; PubMed Scopus Google Scholar). This is by cell neurons of as with cell that M. M. J. 2000; PubMed Scopus Google Scholar). H. Hattori N. Kubo S. M. T. Matsumine H. Asakawa S. Minoshima S. Yamamura Y. Shimizu N. Mizuno Y. PubMed Scopus Google the of an of in brain from patients with is in substantia with antibodies are to a parkin cleavage with a proteolysis in of substantia nigra not The of putative in cleavage of parkin the of antibodies that parkin as protein D. Y. Huang J. A. D. M. H. S. Cell. 97: Full Text Full Text PDF PubMed Scopus Google Scholar). Parkin cleavage to a in neurons caspase activation, by in Parkinson's parkin function that the cellular and to further caspase the neurons a is a demonstrated in a of of the of the protein was by of the symptoms of the A. R. Cell. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). not of the mutation as with parkin the with staurosporine, and not the of an as the of parkin demonstrated to rather parkin cleavage a in Parkinson's disease the of dopaminergic neurons that the of of used in Parkinson's are to a as by the large cell loss M. P. H. P. Nat. Med. PubMed Scopus Google Scholar). of parkin Asp-126 in an for the survival the mutation not the activity of Mizuno and Hattori for the of the human parkin and for of the and for