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Title: The mechanism of β- N -methylamino-l-alanine inhibition of tRNA aminoacylation and its impact on misincorporation
β- N -methylamino- l -alanine (BMAA) is a nonproteinogenic amino acid that has been associated with neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS) and Alzheimer's disease (AD). BMAA has been found in human protein extracts; however, the mechanism by which it enters the proteome is still unclear. It has been suggested that BMAA is misincorporated at serine codons during protein synthesis, but direct evidence of its cotranslational incorporation is currently lacking. Here, using LC-MS–purified BMAA and several biochemical assays, we sought to determine whether any aminoacyl-tRNA synthetase (aaRS) utilizes BMAA as a substrate for aminoacylation. Despite BMAA's previously predicted misincorporation at serine codons, following a screen for amino acid activation in ATP/PP i exchange assays, we observed that BMAA is not a substrate for human seryl-tRNA synthetase (SerRS). Instead, we observed that BMAA is a substrate for human alanyl-tRNA synthetase (AlaRS) and can form BMAA-tRNA Ala by escaping from the intrinsic AlaRS proofreading activity. Furthermore, we found that BMAA inhibits both the cognate amino acid activation and the editing functions of AlaRS. Our results reveal that, in addition to being misincorporated during translation, BMAA may be able to disrupt the integrity of protein synthesis through multiple different mechanisms.  more » « less
Award ID(s):
1715840 2101998
NSF-PAR ID:
10163495
Author(s) / Creator(s):
; ; ; ; ; ;
Date Published:
Journal Name:
Journal of Biological Chemistry
Volume:
295
Issue:
5
ISSN:
0021-9258
Page Range / eLocation ID:
1402 to 1410
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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