In:
PROTEOMICS, Wiley, Vol. 13, No. 9 ( 2013-05), p. 1519-1527
Abstract:
The aspartyl protease BACE 1 cleaves neuregulin 1 and is involved in myelination and is a candidate drug target for A lzheimer's disease, where it acts as the β‐secretase cleaving the amyloid precursor protein. However, little is known about other substrates in vivo. Here, we provide a proteomic workflow for BACE 1 substrate identification from whole brains, combining filter‐aided sample preparation, strong‐anion exchange fractionation, and label‐free quantification. We used bace1 ‐deficient zebrafish and quantified differences in protein levels between wild‐type and bace1 −/− zebrafish brains. Over 4500 proteins were identified with at least two unique peptides and quantified in both wild‐type and bace1 −/− zebrafish brains. The majority of zebrafish membrane proteins did not show altered protein levels, indicating that B ace1 has a restricted substrate specificity. Twenty‐four membrane proteins accumulated in the bace1 −/− brains and thus represent candidate B ace1 substrates. They include several known BACE 1 substrates, such as the zebrafish homologs of amyloid precursor protein and the cell adhesion protein L 1, which validate the proteomic workflow. Additionally, several candidate substrates with a function in neurite outgrowth and axon guidance, such as plexin A 3 and glypican‐1 were identified, pointing to a function of B ace1 in neurodevelopment. Taken together, our study provides the first proteomic analysis of knock‐out zebrafish tissue and demonstrates that combining gene knock‐out models in zebrafish with quantitative proteomics is a powerful approach to address biomedical questions.
Type of Medium:
Online Resource
ISSN:
1615-9853
,
1615-9861
DOI:
10.1002/pmic.201200582
Language:
English
Publisher:
Wiley
Publication Date:
2013
detail.hit.zdb_id:
2037674-1
SSG:
12
Permalink