The neural cell adhesion molecules L1 and CHL1 are cleaved by BACE1 protease in vivo.
The β-site amyloid precursor protein-cleaving enzyme BACE1 is a prime drug target for Alzheimer disease. However, the function and the physiological substrates of BACE1 remain largely unknown. In this work, we took a quantitative proteomic approach to analyze the secretome of primary neurons after acute BACE1 inhibition, and we identified several novel substrate candidates for BACE1. Many of these molecules are involved in neuronal network formation in the developing nervous system. We selected the adhesion molecules L1 and CHL1, which are crucial for axonal guidance and maintenance of neural circuits, for further validation as BACE1 substrates. Using both genetic BACE1 knock-out and acute pharmacological BACE1 inhibition in mice and cell cultures, we show that L1 and CHL1 are cleaved by BACE1 under physiological conditions. The BACE1 cleavage sites at the membrane-proximal regions of L1 (between Tyr(1086) and Glu(1087)) and CHL1 (between Gln(1061) and Asp(1062)) were determined by mass spectrometry. This work provides molecular insights into the function and the pathways in which BACE1 is involved, and it will help to predict or interpret possible side effects of BACE1 inhibitor drugs in current clinical trials.
VIB Center for the Biology of Disease, KULeuven, 3000 Leuven, Belgium.
Lujia
L
Zhou
Soraia
S
Barão
Mathias
M
Laga
Katrijn
K
Bockstael
Marianne
M
Borgers
Harry
H
Gijsen
Wim
W
Annaert
Diederik
D
Moechars
Marc
M
Mercken
Kris
K
Gevaert
Kris
K
Gevaer
Bart
B
De Strooper
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D001921:Brain / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D019556:COS Cells; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D039842:Neural Cell Adhesion Molecule L1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000201:enzymology / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry; D061251:Primary Cell Culture; D011480:Protease Inhibitors / Q000494:pharmacology; D059748:Proteolysis; D020543:Proteome / Q000378:metabolism; D013569:Synapses / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism
D015815:Cell Adhesion Molecules; C490649:Chl1 protein, mouse; D039842:Neural Cell Adhesion Molecule L1; D010446:Peptide Fragments; D011480:Protease Inhibitors; D020543:Proteome; D053829:Amyloid Precursor Protein Secretases; D016282:Aspartic Acid Endopeptidases; C509282:Bace1 protein, mouse