{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T17:20:45Z","timestamp":1791048045384,"version":"4.1.0"},"reference-count":39,"publisher":"National Academy of Sciences","issue":"6","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[1999,3,16]]},"abstract":"<jats:p>\n                    Proteolytic processing of the amyloid precursor protein by \u03b2-secretase yields A4CT (C99), which is cleaved further by the as yet unknown \u03b3-secretase, yielding the \u03b2-amyloid (A\u03b2) peptide with 40 (A\u03b2\n                    <jats:sub>40<\/jats:sub>\n                    ) or 42 residues (A\u03b2\n                    <jats:sub>42<\/jats:sub>\n                    ). Because the position of \u03b3-secretase cleavage is crucial for the pathogenesis of Alzheimer\u2019s disease, we individually replaced all membrane-domain residues of A4CT outside the A\u03b2 domain with phenylalanine, stably transfected the constructs in COS7 cells, and determined the effect of these mutations on the cleavage specificity of \u03b3-secretase (A\u03b2\n                    <jats:sub>42<\/jats:sub>\n                    \/A\u03b2\n                    <jats:sub>40<\/jats:sub>\n                    ratio). Compared with wild-type A4CT, mutations at Val-44, Ile-47, and Val-50 led to decreased A\u03b2\n                    <jats:sub>42<\/jats:sub>\n                    \/A\u03b2\n                    <jats:sub>40<\/jats:sub>\n                    ratios, whereas mutations at Thr-43, Ile-45, Val-46, Leu-49, and Met-51 led to increased A\u03b2\n                    <jats:sub>42<\/jats:sub>\n                    \/A\u03b2\n                    <jats:sub>40<\/jats:sub>\n                    ratios. A massive effect was observed for I45F (34-fold increase) making this construct important for the generation of animal models for Alzheimer\u2019s disease. Unlike the other mutations, A4CT-V44F was processed mainly to A\u03b2\n                    <jats:sub>38<\/jats:sub>\n                    , as determined by mass spectrometry. Our data provide a detailed model for the active site of \u03b3-secretase: \u03b3-secretase interacts with A4CT by binding to one side of the \u03b1-helical transmembrane domain of A4CT. Mutations in the transmembrane domain of A4CT interfere with the interaction between \u03b3-secretase and A4CT and, thus, alter the cleavage specificity of \u03b3-secretase.\n                  <\/jats:p>","DOI":"10.1073\/pnas.96.6.3053","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T10:35:07Z","timestamp":1027679707000},"page":"3053-3058","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":203,"title":["Mechanism of the cleavage specificity of Alzheimer\u2019s disease \u03b3-secretase identified by phenylalanine-scanning mutagenesis of the transmembrane domain of the amyloid precursor protein"],"prefix":"10.1073","volume":"96","author":[{"given":"Stefan F.","family":"Lichtenthaler","sequence":"first","affiliation":[{"name":"Center for Molecular Biology, University of Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Laboratory for Mass Spectrometry, The Rockefeller University, New York, NY 10021; and Department of Pathology, University of Melbourne, Parkville, Victoria 3052, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rong","family":"Wang","sequence":"additional","affiliation":[{"name":"Center for Molecular Biology, University of Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Laboratory for Mass Spectrometry, The Rockefeller University, New York, NY 10021; and Department of Pathology, University of Melbourne, Parkville, Victoria 3052, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heike","family":"Grimm","sequence":"additional","affiliation":[{"name":"Center for Molecular Biology, University of Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Laboratory for Mass Spectrometry, The Rockefeller University, New York, NY 10021; and Department of Pathology, University of Melbourne, Parkville, Victoria 3052, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sacha N.","family":"Uljon","sequence":"additional","affiliation":[{"name":"Center for Molecular Biology, University of Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Laboratory for Mass Spectrometry, The Rockefeller University, New York, NY 10021; and Department of Pathology, University of Melbourne, Parkville, Victoria 3052, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Colin L.","family":"Masters","sequence":"additional","affiliation":[{"name":"Center for Molecular Biology, University of Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Laboratory for Mass Spectrometry, The Rockefeller University, New York, NY 10021; and Department of Pathology, University of Melbourne, Parkville, Victoria 3052, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Konrad","family":"Beyreuther","sequence":"additional","affiliation":[{"name":"Center for Molecular Biology, University of Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Laboratory for Mass Spectrometry, The Rockefeller University, New York, NY 10021; and Department of Pathology, University of Melbourne, Parkville, Victoria 3052, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[1999,3,16]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-291X(84)80190-4"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.82.12.4245"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/325733a0"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.3109\/13506129409146118"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0962-8924(98)01363-4"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.5.2092"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/0896-6273(95)90322-4"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/359322a0"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/359325a0"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1439760"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.93.23.13170"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm0997-1021"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm0997-1016"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.8.4125"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.26.16085"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.141.4.1031"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.50.31894"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.8191290"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1996.1577"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi971071m"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.45.28655"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi970209y"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1021\/jm970621b"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(93)80446-2"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.37.22908"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(87)90587-2"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/6.12.2087"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(93)90312-E"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00069a001"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.38.22999"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.12.5416"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1993.1066"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.36.21914"},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-291X(67)80055-X"},{"key":"e_1_3_3_35_2","doi-asserted-by":"publisher","DOI":"10.1038\/34910"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1097-2765(00)80006-4"},{"key":"e_1_3_3_37_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1996.tb00796.x"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0166-2236(97)01093-X"},{"key":"e_1_3_3_39_2","first-page":"717","volume-title":"Alzheimer\u2019s Disease: Biology, Diagnosis and Therapeutics","author":"Allsop D","year":"1997","unstructured":"D Allsop, G Christie, C Gray, S Holmes, R Markwell, D Owen, L Smith, H Wadsworth, R V Ward, T Hartmann, et al. Alzheimer\u2019s Disease: Biology, Diagnosis and Therapeutics, eds K Iqbal, B Winblad, T Nishimura, M Takeda, H M Wisniewski (Wiley, New York), pp. 717\u2013727 (1997)."}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.96.6.3053","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,13]],"date-time":"2022-04-13T17:51:34Z","timestamp":1649872294000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.96.6.3053"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1999,3,16]]},"references-count":39,"journal-issue":{"issue":"6","published-print":{"date-parts":[[1999,3,16]]}},"alternative-id":["10.1073\/pnas.96.6.3053"],"URL":"https:\/\/doi.org\/10.1073\/pnas.96.6.3053","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[1999,3,16]]},"assertion":[{"value":"1998-09-21","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"1999-03-16","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}