Functional γ-secretase complex assembly in Golgi/trans-Golgi network: interactions among presenilin, nicastrin, Aph1, Pen-2, and γ-secretase substrates
γ-Secretase is a proteolytic complex whose substrates include Notch, β-amyloid precursor protein (APP), and several other type I transmembrane proteins. Presenilin (PS) and nicastrin are known components of this high-molecular-weight complex, and recent genetic screens in invertebrates have identifi...
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| Format: | Article |
| Language: | English |
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Elsevier
2003-11-01
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| Series: | Neurobiology of Disease |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0969996103001232 |
| _version_ | 1818414558096654336 |
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| author | Stephanie Baulac Matthew J LaVoie W.Taylor Kimberly Jennifer Strahle Michael S Wolfe Dennis J Selkoe Weiming Xia |
| author_facet | Stephanie Baulac Matthew J LaVoie W.Taylor Kimberly Jennifer Strahle Michael S Wolfe Dennis J Selkoe Weiming Xia |
| author_sort | Stephanie Baulac |
| collection | DOAJ |
| description | γ-Secretase is a proteolytic complex whose substrates include Notch, β-amyloid precursor protein (APP), and several other type I transmembrane proteins. Presenilin (PS) and nicastrin are known components of this high-molecular-weight complex, and recent genetic screens in invertebrates have identified two additional gene products, Aph1 and Pen-2, as key factors in γ-secretase activity. Here, we examined the interaction of the components of the γ-secretase complex in Chinese hamster ovary cells stably expressing human forms of APP, PS1, Aph1, and Pen-2. Subcellular fractionation of membrane vesicles and subsequent coimmunoprecipitation of individual γ-secretase components revealed that interactions among all proteins occurred in the Golgi/trans-Golgi network (TGN) compartments. Furthermore, incubation of the Golgi/TGN-enriched vesicles resulted in de novo generation of amyloid β-protein and APP intracellular domain. Immunofluorescent staining of the individual γ-secretase components supported our biochemical evidence that the γ-secretase components assemble into the proteolytically active γ-secretase complex in the Golgi/TGN compartment. |
| first_indexed | 2024-12-14T11:21:00Z |
| format | Article |
| id | doaj.art-785261d72ba441759d186202e3adcd89 |
| institution | Directory Open Access Journal |
| issn | 1095-953X |
| language | English |
| last_indexed | 2024-12-14T11:21:00Z |
| publishDate | 2003-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neurobiology of Disease |
| spelling | doaj.art-785261d72ba441759d186202e3adcd892022-12-21T23:03:46ZengElsevierNeurobiology of Disease1095-953X2003-11-01142194204Functional γ-secretase complex assembly in Golgi/trans-Golgi network: interactions among presenilin, nicastrin, Aph1, Pen-2, and γ-secretase substratesStephanie Baulac0Matthew J LaVoie1W.Taylor Kimberly2Jennifer Strahle3Michael S Wolfe4Dennis J Selkoe5Weiming Xia6Center for Neurologic Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USACenter for Neurologic Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USACenter for Neurologic Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USACenter for Neurologic Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USACenter for Neurologic Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USACenter for Neurologic Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USACenter for Neurologic Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USAγ-Secretase is a proteolytic complex whose substrates include Notch, β-amyloid precursor protein (APP), and several other type I transmembrane proteins. Presenilin (PS) and nicastrin are known components of this high-molecular-weight complex, and recent genetic screens in invertebrates have identified two additional gene products, Aph1 and Pen-2, as key factors in γ-secretase activity. Here, we examined the interaction of the components of the γ-secretase complex in Chinese hamster ovary cells stably expressing human forms of APP, PS1, Aph1, and Pen-2. Subcellular fractionation of membrane vesicles and subsequent coimmunoprecipitation of individual γ-secretase components revealed that interactions among all proteins occurred in the Golgi/trans-Golgi network (TGN) compartments. Furthermore, incubation of the Golgi/TGN-enriched vesicles resulted in de novo generation of amyloid β-protein and APP intracellular domain. Immunofluorescent staining of the individual γ-secretase components supported our biochemical evidence that the γ-secretase components assemble into the proteolytically active γ-secretase complex in the Golgi/TGN compartment.http://www.sciencedirect.com/science/article/pii/S0969996103001232AlzheimerPresenilinAmyloidSecretaseAph1Pen-2 |
| spellingShingle | Stephanie Baulac Matthew J LaVoie W.Taylor Kimberly Jennifer Strahle Michael S Wolfe Dennis J Selkoe Weiming Xia Functional γ-secretase complex assembly in Golgi/trans-Golgi network: interactions among presenilin, nicastrin, Aph1, Pen-2, and γ-secretase substrates Neurobiology of Disease Alzheimer Presenilin Amyloid Secretase Aph1 Pen-2 |
| title | Functional γ-secretase complex assembly in Golgi/trans-Golgi network: interactions among presenilin, nicastrin, Aph1, Pen-2, and γ-secretase substrates |
| title_full | Functional γ-secretase complex assembly in Golgi/trans-Golgi network: interactions among presenilin, nicastrin, Aph1, Pen-2, and γ-secretase substrates |
| title_fullStr | Functional γ-secretase complex assembly in Golgi/trans-Golgi network: interactions among presenilin, nicastrin, Aph1, Pen-2, and γ-secretase substrates |
| title_full_unstemmed | Functional γ-secretase complex assembly in Golgi/trans-Golgi network: interactions among presenilin, nicastrin, Aph1, Pen-2, and γ-secretase substrates |
| title_short | Functional γ-secretase complex assembly in Golgi/trans-Golgi network: interactions among presenilin, nicastrin, Aph1, Pen-2, and γ-secretase substrates |
| title_sort | functional γ secretase complex assembly in golgi trans golgi network interactions among presenilin nicastrin aph1 pen 2 and γ secretase substrates |
| topic | Alzheimer Presenilin Amyloid Secretase Aph1 Pen-2 |
| url | http://www.sciencedirect.com/science/article/pii/S0969996103001232 |
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