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Protein Sci. 2017 Aug;26(8):1505-1516. doi: 10.1002/pro.3176. Epub 2017 May 3.

Kinetic and structural changes in HsmtPheRS, induced by pathogenic mutations in human FARS2.

Author information

1
Department of Structural Biology, Weizmann Institute of Science, Israel.
2
McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins School of Medicine, Baltimore.
3
Laboratory of Bioorganic Chemistry of Enzymes, Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia.
4
Ambry Genetics, California.
5
Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas.
6
Wellcome Centre for Mitochondrial Research, Newcastle University, Newcastle, NE2 4HH, United Kingdom.

Abstract

Mutations in the mitochondrial aminoacyl-tRNA synthetases (mtaaRSs) can cause profound clinical presentations, and have manifested as diseases with very selective tissue specificity. To date most of the mtaaRS mutations could be phenotypically recognized, such that clinicians could identify the affected mtaaRS from the symptoms alone. Among the recently reported pathogenic variants are point mutations in FARS2 gene, encoding the human mitochondrial PheRS. Patient symptoms range from spastic paraplegia to fatal infantile Alpers encephalopathy. How clinical manifestations of these mutations relate to the changes in three-dimensional structures and kinetic characteristics remains unclear, although impaired aminoacylation has been proposed as possible etiology of diseases. Here, we report four crystal structures of HsmtPheRS mutants, and extensive MD simulations for wild-type and nine mutants to reveal the structural changes on dynamic trajectories of HsmtPheRS. Using steady-state kinetic measurements of phenylalanine activation and tRNAPhe aminoacylation, we gained insight into the structural and kinetic effects of mitochondrial disease-related mutations in FARS2 gene.

KEYWORDS:

X-ray structures; kinetic experiments; mitochondrial PheRS; mitochondrial diseases; molecular dynamic simulations; mutants

PMID:
28419689
PMCID:
PMC5521548
DOI:
10.1002/pro.3176
[Indexed for MEDLINE]
Free PMC Article

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