Interaction between presenilin 1 and ubiquilin 1 as detected by fluorescence lifetime imaging microscopy and a high-throughput fluorescent plate reader.
Presenilin 1 (PS1) in its active heterodimeric form is the catalytic center of the gamma-secretase complex, an enzymatic activity that cleaves amyloid precursor protein (APP) to produce amyloid beta (Abeta). Ubiquilin 1 is a recently described PS1 interacting protein, the overexpression of which increases PS1 holoprotein levels and leads to reduced levels of functionally active PS1 heterodimer. In addition, it has been suggested that splice variants of the UBQLN1 gene are associated with an increased risk of developing Alzheimer disease (AD). However, it is still unclear whether PS1 and ubiquilin 1 interact when expressed at endogenous levels under normal physiological conditions. Here, we employ three novel fluorescence resonance energy transfer-based techniques to investigate the interaction between PS1 and ubiquilin 1 in intact cells. We consistently find that the ubiquilin 1 N terminus is in close proximity to several epitopes on PS1. We show that ubiquilin 1 interacts both with PS1 holoprotein and heterodimer and that the interaction between PS1 and ubiquilin 1 takes place near the cell surface. Furthermore, we show that the PS1-ubiquilin 1 interaction can be detected between endogenous proteins in primary neurons in vitro as well as in brain tissue of healthy controls and Alzheimer disease patients, providing evidence of its physiological relevance.
Thomas
Anne V
AV
Alzheimer's Disease Research Laboratory, Harvard Medical School, Charlestown, Massachusetts 02129, USA.
Herl
Lauren
L
Spoelgen
Robert
R
Hiltunen
Mikko
M
Jones
Phill B
PB
Tanzi
Rudolph E
RE
Hyman
Bradley T
BT
Berezovska
Oksana
O
D048868:Adaptor Proteins, Signal Transducing; D000368:Aged; D000369:Aged, 80 and over; D000544:Alzheimer Disease / Q000378:metabolism; D000818:Animals; D000071183:Autophagy-Related Proteins; D001921:Brain / Q000166:cytology / Q000473:pathology; D016466:CHO Cells; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D006224:Cricetinae; D019281:Dimerization; D000939:Epitopes; D005260:Female; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D008297:Male; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D008875:Middle Aged; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D053764:Presenilin-1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation
D048868:Adaptor Proteins, Signal Transducing; D000071183:Autophagy-Related Proteins; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D000939:Epitopes; D053764:Presenilin-1; C107792:UBQLN1 protein, human