Phosphorylation Regulates Intracellular Trafficking of β-Secretase

β-Secretase (BACE) is a transmembrane aspartyl protease, which generates the N terminus of Alzheimer's disease amyloid β-peptide. Here, we report that BACE can be phosphorylated within its cytoplasmic domain at serine residue 498 by casein kinase 1. Phosphorylation exclusively occurs after full maturation of BACE by propeptide cleavage and complex N-glycosylation. Phosphorylation/dephosphorylation affects the subcellular localization of BACE. BACE wild type and an S498D mutant that mimics phosphorylated BACE are predominantly located within juxtanuclear Golgi compartments and endosomes, whereas nonphosphorylatable BACE S498A accumulates in peripheral EEA1-positive endosomes. Antibody uptake assays revealed that reinternalization of BACE from the cell surface is independent of its phosphorylation state. After reinternalization, BACE wild type as well as BACE S498D are efficiently retrieved from early endosomal compartments and further targeted to later endosomal compartments and/or the trans-Golgi network. In contrast, nonphosphorylatable BACE S498A is retained within early endosomes. Our results therefore demonstrate regulated trafficking of BACE within the secretory and endocytic pathway. β-Secretase (BACE) is a transmembrane aspartyl protease, which generates the N terminus of Alzheimer's disease amyloid β-peptide. Here, we report that BACE can be phosphorylated within its cytoplasmic domain at serine residue 498 by casein kinase 1. Phosphorylation exclusively occurs after full maturation of BACE by propeptide cleavage and complex N-glycosylation. Phosphorylation/dephosphorylation affects the subcellular localization of BACE. BACE wild type and an S498D mutant that mimics phosphorylated BACE are predominantly located within juxtanuclear Golgi compartments and endosomes, whereas nonphosphorylatable BACE S498A accumulates in peripheral EEA1-positive endosomes. Antibody uptake assays revealed that reinternalization of BACE from the cell surface is independent of its phosphorylation state. After reinternalization, BACE wild type as well as BACE S498D are efficiently retrieved from early endosomal compartments and further targeted to later endosomal compartments and/or the trans-Golgi network. In contrast, nonphosphorylatable BACE S498A is retained within early endosomes. Our results therefore demonstrate regulated trafficking of BACE within the secretory and endocytic pathway. amyloid β-peptide brefeldin A β-amyloid precursor protein casein kinase cytoplasmic tail early endosome antigen 1 glutathione S-transferase hymenialdisine human embryonic kidney polyvinylidene difluoride polyacrylamide gel electrophoresis β-secretase endoglycosidase H phosphate-buffered saline endoplasmic reticulum trans-Golgi network wild type peptide:N-glycanase F. Alzheimer's disease is the most common form of dementia and is pathologically characterized by the invariant accumulation of senile plaques and neurofibrillary tangles in certain areas of the brains of Alzheimer's disease patients (1Selkoe D.J. Nature. 1999; 399: A23-A31Crossref PubMed Scopus (1519) Google Scholar). The major constituent of senile plaques is the amyloid β-peptide (Aβ),1 which derives from the β-amyloid precursor protein (βAPP) by endoproteolytic processing (2Haass C. Selkoe D.J. Cell. 1993; 75: 1039-1042Abstract Full Text PDF PubMed Scopus (736) Google Scholar). β-Secretase cleaves βAPP at the N terminus of the Aβ-domain, resulting in the generation of soluble APPS-β and a membrane-associated C-terminal fragment bearing the complete Aβ-domain. Subsequent cleavage of this fragment by γ-secretase, which appears to be identical with the presenilins, results in the release and secretion of Aβ (3Haass C. De Strooper B. Science. 1999; 286: 916-919Crossref PubMed Scopus (364) Google Scholar).Recently, an aspartyl protease with β-secretase activity was identified in human embryonic kidney (HEK) 293 cells and was initially called BACE (β-site APP-cleaving enzyme, Asp2, or memapsin 2) (4Hussain I. Powell D. Howlett D.R. Tew D.G. Meek T.D. Chapman C. Gloger I.S. Murphy K.E. Southan C.D. Ryan D.M. Smith T.S. Simmons D.L. Walsh F.S. Dingwall C. Christie G. Mol. Cell Neurosci. 1999; 14: 419-427Crossref PubMed Scopus (997) Google Scholar, 5Lin X. Koelsch G. Wu S. Downs D. Dashti A. Tang J. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 1456-1460Crossref PubMed Scopus (737) Google Scholar, 6Sinha S. Anderson J.P. Barbour R. Basi G.S. Caccavello R. Davis D. Doan M. Dovey H.F. Frigon N. Hong J. Jacobson-Croak K. Jewett N. Keim P. Knops J. Lieberburg I. Power M. Tan H. Tatsuno G. Tung J. Schenk D. Seubert P. Suomensaari S.M. Wang S. Walker D. John V. et al.Nature. 1999; 402: 537-540Crossref PubMed Scopus (1472) Google Scholar, 7Vassar R. Bennett B.D. Babu-Khan S. Kahn S. Mendiaz E.A. Denis P. Teplow D.B. Ross S. Amarante P. Loeloff R. Luo S. J. S. J. F. J. G. M. Science. 1999; 286: PubMed Scopus Google Scholar, R. H. D.B. Nature. 1999; 402: PubMed Scopus Google 1 A was identified and as or memapsin 1 (4Hussain I. Powell D. Howlett D.R. Tew D.G. Meek T.D. Chapman C. Gloger I.S. Murphy K.E. Southan C.D. Ryan D.M. Smith T.S. Simmons D.L. Walsh F.S. Dingwall C. Christie G. Mol. Cell Neurosci. 1999; 14: 419-427Crossref PubMed Scopus (997) Google Scholar, 5Lin X. Koelsch G. Wu S. Downs D. Dashti A. Tang J. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 1456-1460Crossref PubMed Scopus (737) Google F. M. C. P. S. R. 2000; PubMed Scopus Google Scholar). of we the BACE and in this of we the BACE and in this are type with of the aspartyl protease X. Koelsch G. Wu S. Downs D. Dashti A. Tang J. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 1456-1460Crossref PubMed Scopus (737) Google Scholar, 6Sinha S. Anderson J.P. Barbour R. Basi G.S. Caccavello R. Davis D. Doan M. Dovey H.F. Frigon N. Hong J. Jacobson-Croak K. Jewett N. Keim P. Knops J. Lieberburg I. Power M. Tan H. Tatsuno G. Tung J. Schenk D. Seubert P. Suomensaari S.M. Wang S. Walker D. John V. et al.Nature. 1999; 402: 537-540Crossref PubMed Scopus (1472) Google Scholar, 7Vassar R. Bennett B.D. Babu-Khan S. Kahn S. Mendiaz E.A. Denis P. Teplow D.B. Ross S. Amarante P. Loeloff R. Luo S. J. S. J. F. J. G. M. Science. 1999; 286: PubMed Scopus Google Scholar, R. H. D.B. Nature. 1999; 402: PubMed Scopus Google Scholar). is predominantly in peripheral BACE is in the major of Aβ appears that βAPP is the BACE. In βAPP is a β-secretase activity Strooper B. M. G. F. K. J. 14: PubMed Scopus Google Scholar). the Aβ domain of βAPP by in cleavage by β-secretase Strooper B. M. G. F. K. J. 14: PubMed Scopus Google that βAPP be the BACE. of BACE to be is by and further by complex as well as of its by a protease B.D. Denis P. M. Teplow D.B. Kahn S. M. R. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, G. K. De Strooper B. J. Full Text Full Text PDF PubMed Scopus Google Scholar, J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). The of BACE are within Golgi and endosomal with βAPP (4Hussain I. Powell D. Howlett D.R. Tew D.G. Meek T.D. Chapman C. Gloger I.S. Murphy K.E. Southan C.D. Ryan D.M. Smith T.S. Simmons D.L. Walsh F.S. Dingwall C. Christie G. Mol. Cell Neurosci. 1999; 14: 419-427Crossref PubMed Scopus (997) Google Scholar, 5Lin X. Koelsch G. Wu S. Downs D. Dashti A. Tang J. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 1456-1460Crossref PubMed Scopus (737) Google Scholar, 7Vassar R. Bennett B.D. Babu-Khan S. Kahn S. Mendiaz E.A. Denis P. Teplow D.B. Ross S. Amarante P. Loeloff R. Luo S. J. S. J. F. J. G. M. Science. 1999; 286: PubMed Scopus Google Scholar, A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). The of BACE X. Koelsch G. Wu S. Downs D. Dashti A. Tang J. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 1456-1460Crossref PubMed Scopus (737) Google Scholar, 6Sinha S. Anderson J.P. Barbour R. Basi G.S. Caccavello R. Davis D. Doan M. Dovey H.F. Frigon N. Hong J. Jacobson-Croak K. Jewett N. Keim P. Knops J. Lieberburg I. Power M. Tan H. Tatsuno G. Tung J. Schenk D. Seubert P. Suomensaari S.M. Wang S. Walker D. John V. et al.Nature. 1999; 402: 537-540Crossref PubMed Scopus (1472) Google Scholar, 7Vassar R. Bennett B.D. Babu-Khan S. Kahn S. Mendiaz E.A. Denis P. Teplow D.B. Ross S. Amarante P. Loeloff R. Luo S. J. S. J. F. J. G. M. Science. 1999; 286: PubMed Scopus Google Scholar, R. H. D.B. Nature. 1999; 402: PubMed Scopus Google that is predominantly within Golgi compartments and/or is with that β-secretase cleavage of βAPP can in of compartments C. A. Selkoe D.J. Nature. PubMed Scopus Google Scholar, C. A. M. Seubert P. Schenk D. Selkoe D.J. PubMed Scopus Google Scholar, J. Full Text PDF PubMed Google Scholar, Selkoe D.J. J. Cell Sci. PubMed Google that BACE is from the cell surface to early and can to the cell a that a in the cytoplasmic tail of BACE J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). is located to a which a phosphorylation that BACE is phosphorylated within its C-terminal domain and that the of BACE phosphorylation in the of the of BACE from this we the subcellular trafficking of BACE its phosphorylation state. A phosphorylation was identified by of serine residue 498 in the C-terminal domain of BACE 1 protein we that can BACE at phosphorylation at serine residue which kinase and M. M. J. K.E. Wu J. 2000; Full Text Full Text PDF PubMed Scopus Google phosphorylation of the cytoplasmic domain of BACE in The phosphorylation identified at serine residue 498 is by a of 1 a kinase or that or a kinase is in the phosphorylation of BACE in Phosphorylation occurs exclusively the BACE after propeptide and that phosphorylation of BACE after its from the in Golgi or occurs in and to be with the and and to a of Anderson Cell PubMed Scopus Google Scholar). is in the of trafficking in by of Mol. Cell. 2000; PubMed Scopus Google Scholar, Tan J. PubMed Scopus Google Scholar). Our that a with a subcellular in Golgi and/or endosomal be the phosphorylation of BACE. that is in Alzheimer's disease brains C. N. J. 2000; PubMed Scopus Google Scholar, K. J. 2000; PubMed Scopus Google Scholar). Aβ to in A. N. H. 1993; PubMed Scopus Google Scholar). to be phosphorylation of BACE is of Alzheimer's to the of the trafficking of we in which the phosphorylation at serine residue 498 by an or an residue to and phosphorylated of The major of this is to the of kinase or that or Cell. 1993; 75: Full Text PDF PubMed Scopus Google Scholar, J. A. J. C. Proc. Natl. Acad. Sci. U. S. A. 1999; PubMed Scopus Google Scholar). BACE the S498D was predominantly in juxtanuclear with the Golgi protein A was BACE which is with results R. Bennett B.D. Babu-Khan S. Kahn S. Mendiaz E.A. Denis P. Teplow D.B. Ross S. Amarante P. Loeloff R. Luo S. J. S. J. F. J. G. M. Science. 1999; 286: PubMed Scopus Google Scholar, A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). In contrast, the form BACE S498A localization in juxtanuclear localization in compartments early endosomes. was that the complete of the cytoplasmic domain of BACE results in its within the and in maturation A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). the phosphorylation BACE S498A and BACE S498D by complex and of the as with BACE the in subcellular localization of the mutant of BACE are to with results A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, J. 2000; Full Text Full Text PDF PubMed Scopus Google we that BACE is to the cell surface and that BACE is EEA1-positive early endosomal BACE as well as the mutant S498A and S498D efficiently that phosphorylation of BACE its reinternalization from the cell we that phosphorylation of BACE is of the from early to later endosomal and/or compartments from which BACE be the secretory phosphorylation of BACE its with its protein βAPP in juxtanuclear BACE or the phosphorylation to secretion of Aβ as with cell that of BACE to of Aβ of the in subcellular localization of the BACE that βAPP be by β-secretase activity in and Golgi compartments C. A. M. Seubert P. Schenk D. Selkoe D.J. PubMed Scopus Google Scholar, J. Full Text PDF PubMed Google Scholar, C. Teplow D.B. J. Full Text PDF PubMed Google Scholar). demonstrate that reinternalization of BACE was by the S498A or S498D In of BACE maturation by complex N-glycosylation. that at major of β-secretase and secretory can be by BACE independent of its phosphorylation state. be that of subcellular trafficking of BACE by phosphorylation affects processing of protein In we the that phosphorylation βAPP of subcellular trafficking of BACE is to that of of from endosomal to compartments was to be of its cytoplasmic domain C. G. J. Cell PubMed Scopus Google Scholar). the subcellular localization of BACE is regulated in a a protease that to be propeptide cleavage of BACE B.D. Denis P. M. Teplow D.B. Kahn S. M. R. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, G. K. De Strooper B. J. Full Text Full Text PDF PubMed Scopus Google Scholar). Alzheimer's disease is the most common form of dementia and is pathologically characterized by the invariant accumulation of senile plaques and neurofibrillary tangles in certain areas of the brains of Alzheimer's disease patients (1Selkoe D.J. Nature. 1999; 399: A23-A31Crossref PubMed Scopus (1519) Google Scholar). The major constituent of senile plaques is the amyloid β-peptide (Aβ),1 which derives from the β-amyloid precursor protein (βAPP) by endoproteolytic processing (2Haass C. Selkoe D.J. Cell. 1993; 75: 1039-1042Abstract Full Text PDF PubMed Scopus (736) Google Scholar). β-Secretase cleaves βAPP at the N terminus of the Aβ-domain, resulting in the generation of soluble APPS-β and a membrane-associated C-terminal fragment bearing the complete Aβ-domain. Subsequent cleavage of this fragment by γ-secretase, which appears to be identical with the presenilins, results in the release and secretion of Aβ (3Haass C. De Strooper B. Science. 1999; 286: 916-919Crossref PubMed Scopus (364) Google Scholar). an aspartyl protease with β-secretase activity was identified in human embryonic kidney (HEK) 293 cells and was initially called BACE (β-site APP-cleaving enzyme, Asp2, or memapsin 2) (4Hussain I. Powell D. Howlett D.R. Tew D.G. Meek T.D. Chapman C. Gloger I.S. Murphy K.E. Southan C.D. Ryan D.M. Smith T.S. Simmons D.L. Walsh F.S. Dingwall C. Christie G. Mol. Cell Neurosci. 1999; 14: 419-427Crossref PubMed Scopus (997) Google Scholar, 5Lin X. Koelsch G. Wu S. Downs D. Dashti A. Tang J. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 1456-1460Crossref PubMed Scopus (737) Google Scholar, 6Sinha S. Anderson J.P. Barbour R. Basi G.S. Caccavello R. Davis D. Doan M. Dovey H.F. Frigon N. Hong J. Jacobson-Croak K. Jewett N. Keim P. Knops J. Lieberburg I. Power M. Tan H. Tatsuno G. Tung J. Schenk D. Seubert P. Suomensaari S.M. Wang S. Walker D. John V. et al.Nature. 1999; 402: 537-540Crossref PubMed Scopus (1472) Google Scholar, 7Vassar R. Bennett B.D. Babu-Khan S. Kahn S. Mendiaz E.A. Denis P. Teplow D.B. Ross S. Amarante P. Loeloff R. Luo S. J. S. J. F. J. G. M. Science. 1999; 286: PubMed Scopus Google Scholar, R. H. D.B. Nature. 1999; 402: PubMed Scopus Google 1 A was identified and as or memapsin 1 (4Hussain I. Powell D. Howlett D.R. Tew D.G. Meek T.D. Chapman C. Gloger I.S. Murphy K.E. Southan C.D. Ryan D.M. Smith T.S. Simmons D.L. Walsh F.S. Dingwall C. Christie G. Mol. Cell Neurosci. 1999; 14: 419-427Crossref PubMed Scopus (997) Google Scholar, 5Lin X. Koelsch G. Wu S. Downs D. Dashti A. Tang J. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 1456-1460Crossref PubMed Scopus (737) Google F. M. C. P. S. R. 2000; PubMed Scopus Google Scholar). of we the BACE and in this of we the BACE and in this are type with of the aspartyl protease X. Koelsch G. Wu S. Downs D. Dashti A. Tang J. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 1456-1460Crossref PubMed Scopus (737) Google Scholar, 6Sinha S. Anderson J.P. Barbour R. Basi G.S. Caccavello R. Davis D. Doan M. Dovey H.F. Frigon N. Hong J. Jacobson-Croak K. Jewett N. Keim P. Knops J. Lieberburg I. Power M. Tan H. Tatsuno G. Tung J. Schenk D. Seubert P. Suomensaari S.M. Wang S. Walker D. John V. et al.Nature. 1999; 402: 537-540Crossref PubMed Scopus (1472) Google Scholar, 7Vassar R. Bennett B.D. Babu-Khan S. Kahn S. Mendiaz E.A. Denis P. Teplow D.B. Ross S. Amarante P. Loeloff R. Luo S. J. S. J. F. J. G. M. Science. 1999; 286: PubMed Scopus Google Scholar, R. H. D.B. Nature. 1999; 402: PubMed Scopus Google Scholar). is predominantly in peripheral BACE is in the major of Aβ appears that βAPP is the BACE. In βAPP is a β-secretase activity Strooper B. M. G. F. K. J. 14: PubMed Scopus Google Scholar). the Aβ domain of βAPP by in cleavage by β-secretase Strooper B. M. G. F. K. J. 14: PubMed Scopus Google that βAPP be the BACE. of BACE to be BACE is by and further by complex as well as of its by a protease B.D. Denis P. M. Teplow D.B. Kahn S. M. R. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, G. K. De Strooper B. J. Full Text Full Text PDF PubMed Scopus Google Scholar, J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). The of BACE are within Golgi and endosomal with βAPP (4Hussain I. Powell D. Howlett D.R. Tew D.G. Meek T.D. Chapman C. Gloger I.S. Murphy K.E. Southan C.D. Ryan D.M. Smith T.S. Simmons D.L. Walsh F.S. Dingwall C. Christie G. Mol. Cell Neurosci. 1999; 14: 419-427Crossref PubMed Scopus (997) Google Scholar, 5Lin X. Koelsch G. Wu S. Downs D. Dashti A. Tang J. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 1456-1460Crossref PubMed Scopus (737) Google Scholar, 7Vassar R. Bennett B.D. Babu-Khan S. Kahn S. Mendiaz E.A. Denis P. Teplow D.B. Ross S. Amarante P. Loeloff R. Luo S. J. S. J. F. J. G. M. Science. 1999; 286: PubMed Scopus Google Scholar, A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). The of BACE X. Koelsch G. Wu S. Downs D. Dashti A. Tang J. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 1456-1460Crossref PubMed Scopus (737) Google Scholar, 6Sinha S. Anderson J.P. Barbour R. Basi G.S. Caccavello R. Davis D. Doan M. Dovey H.F. Frigon N. Hong J. Jacobson-Croak K. Jewett N. Keim P. Knops J. Lieberburg I. Power M. Tan H. Tatsuno G. Tung J. Schenk D. Seubert P. Suomensaari S.M. Wang S. Walker D. John V. et al.Nature. 1999; 402: 537-540Crossref PubMed Scopus (1472) Google Scholar, 7Vassar R. Bennett B.D. Babu-Khan S. Kahn S. Mendiaz E.A. Denis P. Teplow D.B. Ross S. Amarante P. Loeloff R. Luo S. J. S. J. F. J. G. M. Science. 1999; 286: PubMed Scopus Google Scholar, R. H. D.B. Nature. 1999; 402: PubMed Scopus Google that is predominantly within Golgi compartments and/or is with that β-secretase cleavage of βAPP can in of compartments C. A. Selkoe D.J. Nature. PubMed Scopus Google Scholar, C. A. M. Seubert P. Schenk D. Selkoe D.J. PubMed Scopus Google Scholar, J. Full Text PDF PubMed Google Scholar, Selkoe D.J. J. Cell Sci. PubMed Google Scholar). that BACE is from the cell surface to early and can to the cell a that a in the cytoplasmic tail of BACE J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). is located to a which a phosphorylation that BACE is phosphorylated within its C-terminal domain and that the of BACE phosphorylation in the of the of BACE from endosomes. this we the subcellular trafficking of BACE its phosphorylation state. A phosphorylation was identified by of serine residue 498 in the C-terminal domain of BACE 1 protein we that can BACE at phosphorylation at serine residue which kinase and M. M. J. K.E. Wu J. 2000; Full Text Full Text PDF PubMed Scopus Google phosphorylation of the cytoplasmic domain of BACE in The phosphorylation identified at serine residue 498 is by a of 1 a kinase or that or a kinase is in the phosphorylation of BACE in Phosphorylation occurs exclusively the BACE after propeptide and that phosphorylation of BACE after its from the in Golgi or occurs in and to be with the and and to a of Anderson Cell PubMed Scopus Google Scholar). is in the of trafficking in by of Mol. Cell. 2000; PubMed Scopus Google Scholar, Tan J. PubMed Scopus Google Scholar). Our that a with a subcellular in Golgi and/or endosomal be the phosphorylation of BACE. that is in Alzheimer's disease brains C. N. J. 2000; PubMed Scopus Google Scholar, K. J. 2000; PubMed Scopus Google Scholar). Aβ to in A. N. H. 1993; PubMed Scopus Google Scholar). to be phosphorylation of BACE is of Alzheimer's to the of the trafficking of we in which the phosphorylation at serine residue 498 by an or an residue to and phosphorylated of The major of this is to the of kinase or that or Cell. 1993; 75: Full Text PDF PubMed Scopus Google Scholar, J. A. J. C. Proc. Natl. Acad. Sci. U. S. A. 1999; PubMed Scopus Google Scholar). BACE the S498D was predominantly in juxtanuclear with the Golgi protein A was BACE which is with results R. Bennett B.D. Babu-Khan S. Kahn S. Mendiaz E.A. Denis P. Teplow D.B. Ross S. Amarante P. Loeloff R. Luo S. J. S. J. F. J. G. M. Science. 1999; 286: PubMed Scopus Google Scholar, A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). In contrast, the form BACE S498A localization in juxtanuclear localization in compartments early endosomes. was that the complete of the cytoplasmic domain of BACE results in its within the and in maturation A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). the phosphorylation BACE S498A and BACE S498D by complex and of the as with BACE the in subcellular localization of the mutant of BACE are to with results A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, J. 2000; Full Text Full Text PDF PubMed Scopus Google we that BACE is to the cell surface and that BACE is EEA1-positive early endosomal BACE as well as the mutant S498A and S498D efficiently that phosphorylation of BACE its reinternalization from the cell we that phosphorylation of BACE is of the from early to later endosomal and/or compartments from which BACE be the secretory phosphorylation of BACE its with its protein βAPP in juxtanuclear BACE or the phosphorylation to secretion of Aβ as with cell that of BACE to of Aβ of the in subcellular localization of the BACE that βAPP be by β-secretase activity in and Golgi compartments C. A. M. Seubert P. Schenk D. Selkoe D.J. PubMed Scopus Google Scholar, J. Full Text PDF PubMed Google Scholar, C. Teplow D.B. J. Full Text PDF PubMed Google Scholar). demonstrate that reinternalization of BACE was by the S498A or S498D In of BACE maturation by complex N-glycosylation. that at major of β-secretase and secretory can be by BACE independent of its phosphorylation state. be that of subcellular trafficking of BACE by phosphorylation affects processing of protein In we the that phosphorylation βAPP of subcellular trafficking of BACE is to that of of from endosomal to compartments was to be of its cytoplasmic domain C. G. J. Cell PubMed Scopus Google Scholar). the subcellular localization of BACE is regulated in a a protease that to be propeptide cleavage of BACE B.D. Denis P. M. Teplow D.B. Kahn S. M. R. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, G. K. De Strooper B. J. Full Text Full Text PDF PubMed Scopus Google Scholar). In this we the subcellular trafficking of BACE its phosphorylation state. A phosphorylation was identified by of serine residue 498 in the C-terminal domain of BACE 1 protein we that can BACE at phosphorylation at serine residue which kinase and M. M. J. K.E. Wu J. 2000; Full Text Full Text PDF PubMed Scopus Google phosphorylation of the cytoplasmic domain of BACE in The phosphorylation identified at serine residue 498 is by a of 1 a kinase or that or a kinase is in the phosphorylation of BACE in Phosphorylation occurs exclusively the BACE after propeptide and that phosphorylation of BACE after its from the in Golgi or occurs in and to be with the and and to a of Anderson Cell PubMed Scopus Google Scholar). is in the of trafficking in by of Mol. Cell. 2000; PubMed Scopus Google Scholar, Tan J. PubMed Scopus Google Scholar). Our that a with a subcellular in Golgi and/or endosomal be the phosphorylation of BACE. that is in Alzheimer's disease brains C. N. J. 2000; PubMed Scopus Google Scholar, K. J. 2000; PubMed Scopus Google Scholar). Aβ to in A. N. H. 1993; PubMed Scopus Google Scholar). to be phosphorylation of BACE is of Alzheimer's In to the of the trafficking of we in which the phosphorylation at serine residue 498 by an or an residue to and phosphorylated of The major of this is to the of kinase or that or Cell. 1993; 75: Full Text PDF PubMed Scopus Google Scholar, J. A. J. C. Proc. Natl. Acad. Sci. U. S. A. 1999; PubMed Scopus Google Scholar). BACE the S498D was predominantly in juxtanuclear with the Golgi protein A was BACE which is with results R. Bennett B.D. Babu-Khan S. Kahn S. Mendiaz E.A. Denis P. Teplow D.B. Ross S. Amarante P. Loeloff R. Luo S. J. S. J. F. J. G. M. Science. 1999; 286: PubMed Scopus Google Scholar, A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). In contrast, the form BACE S498A localization in juxtanuclear localization in compartments early endosomes. was that the complete of the cytoplasmic domain of BACE results in its within the and in maturation A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). the phosphorylation BACE S498A and BACE S498D by complex and of the as with BACE the in subcellular localization of the mutant of BACE are to with results A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, J. 2000; Full Text Full Text PDF PubMed Scopus Google we that BACE is to the cell surface and that BACE is EEA1-positive early endosomal BACE as well as the mutant S498A and S498D efficiently that phosphorylation of BACE its reinternalization from the cell we that phosphorylation of BACE is of the from early to later endosomal and/or compartments from which BACE be the secretory pathway. phosphorylation of BACE its with its protein βAPP in juxtanuclear BACE or the phosphorylation to secretion of Aβ as with cell that of BACE to of Aβ of the in subcellular localization of the BACE that βAPP be by β-secretase activity in and Golgi compartments C. A. M. Seubert P. Schenk D. Selkoe D.J. PubMed Scopus Google Scholar, J. Full Text PDF PubMed Google Scholar, C. Teplow D.B. J. Full Text PDF PubMed Google Scholar). demonstrate that reinternalization of BACE was by the S498A or S498D In of BACE maturation by complex N-glycosylation. that at major of β-secretase and secretory can be by BACE independent of its phosphorylation state. be that of subcellular trafficking of BACE by phosphorylation affects processing of protein In we the that phosphorylation βAPP The of subcellular trafficking of BACE is to that of of from endosomal to compartments was to be of its cytoplasmic domain C. G. J. Cell PubMed Scopus Google Scholar). the subcellular localization of BACE is regulated in a a protease that to be propeptide cleavage of BACE B.D. Denis P. M. Teplow D.B. Kahn S. M. R. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, A. H. M. H. C. J. S. G. C. J. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, G. K. De Strooper B. J. Full Text Full Text PDF PubMed Scopus Google Scholar). are to and H. and to V. protein kinase A. and G. M. and H. and the and