A novel polymorphism at codon 705 in the amyloid precursor protein (APP) gene found with single stranded conformation polymorphism (SSCP) analysis

Cell Line ◽

Quantitative Proteomics ◽

Protein Processing ◽

Precursor Protein ◽

Amyloid Precursor Protein Processing ◽

App Processing ◽

293 Cells ◽

Line Construction ◽

Lyso-IP is a method that allows for the isolation of lysosomes for proteomics and metabolomics (dx.doi.org/10.17504/protocols.io.bybjpskn; dx.doi.org/10.17504/protocols.io.bx9hpr36). We have developed an analogous approach for purification of early/sorting endosomes (Endo-IP). In addition, we have found that endolysosomal purification via Lyso-IP and Endo-IP can be coupled with a quantitative proteomics workflow to obtain snapshots of Amyloid Precursor Protein (APP) processing to its Aβ products (Park et al. in submission). Here, we describe methods for cell line construction and maintenance of 293 cells with TMEM192-3xHA and 3xFLAG-EEA1, which are used for lysosome and endosome purification, respectively, with the addition of patient mutations to APP promotes processing. Cells with endogenously tagged TMEM192 and stably expressing FLAG-EEA1 are referred to as 293EL cells, for Endo-IP and Lyso-IP. These cells were also prepared in a form that has a deletion of the APP gene (293EL;APP-/-) and the same cells reconstituted with a lentivirus stably expressing APPSw;T700N to allow functional analysis of APP processing.

Economic Fluorescent Labeling of PCR Products for Microsatellite- and Single- Stranded Conformation Polymorphism (SSCP) Analysis

PCR Technology ◽

10.1201/9781420040654-13 ◽

2003 ◽

pp. 101-108

Keyword(s):

Fluorescent Labeling ◽

Sscp Analysis ◽

Single Stranded Conformation Polymorphism ◽

Pcr Products ◽

Conformation Polymorphism

No evidence that common allelic variation in the Amyloid Precursor Protein (APP) gene confers susceptibility to Alzheimer's disease

Human Molecular Genetics ◽

10.1093/hmg/4.5.853 ◽

1995 ◽

Vol 4 (5) ◽

pp. 853-858 ◽

Cited By ~ 11

Author(s):

M. B. Liddell ◽

A. J. Bayer ◽

M. J. Owen

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Allelic Variation ◽

Precursor Protein ◽

Species specificity and developmental patterns of expression of the β amyloid precursor protein (APP) gene in brain, liver and choroid plexus in birds

Comparative Biochemistry and Physiology Part B Comparative Biochemistry ◽

10.1016/0305-0491(92)90018-m ◽

1992 ◽

Vol 101 (3) ◽

pp. 391-398 ◽

Cited By ~ 2

Author(s):

Guo-Fen Tu ◽

Bridget R. Southwell ◽

Gerhard Schreiber

Keyword(s):

Choroid Plexus ◽

Species Specificity ◽

Precursor Protein ◽

Developmental Patterns ◽

Β Amyloid ◽

A neurodevelopmental disorder with a nonsense mutation in the Ox-2 antigen domain of the amyloid precursor protein (APP) gene

Nucleosides Nucleotides & Nucleic Acids ◽

10.1080/15257770.2016.1267361 ◽

2017 ◽

pp. 1-11 ◽

Cited By ~ 3

Author(s):

Khue Vu Nguyen ◽

Karen Leydiker ◽

Raymond Wang ◽

Jose Abdenur ◽

William L. Nyhan

Keyword(s):

Nonsense Mutation ◽

Neurodevelopmental Disorder ◽

Precursor Protein ◽

Evaluation of the Expression of Amyloid Precursor Protein and the Ratio of Secreted Amyloid Beta 42 to Amyloid Beta 40 in SH-SY5Y Cells Stably Transfected with Wild-Type, Single-Mutant and Double-Mutant Forms of the APP Gene for the Study of Alzheimer’s Disease Pathology

Applied Biochemistry and Biotechnology ◽

10.1007/s12010-017-2468-6 ◽

2017 ◽

Vol 183 (3) ◽

pp. 853-866 ◽

Cited By ~ 3

Author(s):

Aslina Pahrudin Arrozi ◽

Siti Nur Syazwani Shukri ◽

Wan Zurinah Wan Ngah ◽

Yasmin Anum Mohd Yusof ◽

Mohd Hanafi Ahmad Damanhuri ◽

...

Keyword(s):

Alzheimer’S Disease ◽

Amyloid Beta ◽

Double Mutant ◽

Precursor Protein ◽

Wild Type ◽

Single Mutant ◽

Amyloid Beta 42 ◽

Mutant Forms ◽

Hereditary Alzheimer disease with a guanine to thymine missense change at position 1924 of the amyloid precursor protein (APP) gene

Neurobiology of Aging ◽

10.1016/0197-4580(92)90384-a ◽

1992 ◽

Vol 13 ◽

pp. S65

Keyword(s):

Alzheimer Disease ◽

Precursor Protein ◽

Missense Change ◽

Novel polymorphisms of the amyloid precursor protein (APP) gene in Chinese/Taiwanese patients with Alzheimer’s disease

Journal of Clinical Neuroscience ◽

10.1016/j.jocn.2008.04.006 ◽

2009 ◽

Vol 16 (2) ◽

pp. 259-263 ◽

Cited By ~ 6

Author(s):

Peterus Thajeb ◽

Pei Wang ◽

Chung-Liang Chien ◽

Rosanne Harrigan

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Precursor Protein ◽

Author response: Genomic mosaicism with increased amyloid precursor protein (APP) gene copy number in single neurons from sporadic Alzheimer's disease brains

10.7554/elife.05116.031 ◽

2015 ◽

Cited By ~ 1

Author(s):

Diane M Bushman ◽

Gwendolyn E Kaeser ◽

Benjamin Siddoway ◽

Jurgen W Westra ◽

Richard R Rivera ◽

...

Keyword(s):

Alzheimer’S Disease ◽

Copy Number ◽

Gene Copy Number ◽

Precursor Protein ◽

Author Response ◽

Gene Copy ◽

Single Neurons ◽

Sporadic Alzheimer’S Disease ◽

Amyloid precursor protein metabolism in fibroblasts from individuals with one, two or three copies of the amyloid precursor protein (APP) gene

Biochemical Journal ◽

10.1042/bj3380777 ◽

1999 ◽

Vol 338 (3) ◽

pp. 777-782 ◽

Cited By ~ 4

Author(s):

Marco RACCHI ◽

Janet A. JOHNSTON ◽

Fiona M. FLOOD ◽

Richard F. COWBURN ◽

Stefano GOVONI

Keyword(s):

Phorbol Ester ◽

Protein Metabolism ◽

Precursor Protein ◽

Chromosome 21 ◽

Human Skin Fibroblasts ◽

Kinase C ◽

App Metabolism ◽

App Gene ◽

Pkc Activation

Protein kinase C (PKC)-activated modulation of amyloid precursor protein (APP) metabolism has been investigated in natural models of altered APP expression due to the presence of one, two or three copies of the APP gene. We show that levels of APP present in human skin fibroblasts strongly influence the effect of PKC activation of soluble APP (sAPP) release. Thus fibroblasts derived from a patient with a deletion in chromosome 21 including the APP locus (Δ21) had lower levels of both APP mRNA and cell-associated APP, and showed an exaggerated phorbol-ester-induced sAPP release, when compared with fibroblasts from control individuals. In contrast, fibroblasts from chromosome 21 trisomic Down's syndrome patients failed to show a concentration-dependent response to phorbol ester treatment. These results suggest that the levels of APP expression can affect the degree of response to PKC-mediated modulation of the metabolism of this protein.