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Purification, crystallization and preliminary X-ray characterization of a human mitochondrial phenylalanyl-tRNA synthetase

Journal Article · · Acta Crystallographica. Section F
;  [1];  [2];  [1];  [3];  [4];  [5];  [1]
  1. Department of Structural Biology, Weizmann Institute of Science, 76100 Rehovot (Israel)
  2. Institute of Chemical Biology and Fundamental Medicine, 630090 Novosibirsk (Russian Federation)
  3. Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, PA 16802 (United States)
  4. Department Chemistry and Biochemistry, University of Colorado, Boulder, CO 80309-0215 (United States)
  5. Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599-3290 (United States)
The expression, purification and crystallization of recombinant human mitochondrial phenylalanyl-tRNA synthetase (mitPheRS) are reported. Diffraction data were collected to 2.2 Å resolution and the mitPheRS structure was solved using the molecular-replacement method. Human monomeric mitochondrial phenylalanyl-tRNA synthetase (mitPheRS) is an enzyme that catalyzes the charging of tRNA with the cognate amino acid phenylalanine. Human mitPheRS is a chimera of the bacterial α-subunit of PheRS and the B8 domain of its β-subunit. Together, the α-subunit and the ‘RNP-domain’ (B8 domain) at the C-terminus form the minimal structural set to construct an enzyme with phenylalanylation activity. The recombinant human mitPheRS was purified to homogeneity and crystallized in complex with phenylalanine and ATP. The crystals diffracted to 2.2 Å resolution and belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 55, b = 90, c = 96 Å.
OSTI ID:
22360529
Journal Information:
Acta Crystallographica. Section F, Journal Name: Acta Crystallographica. Section F Journal Issue: Pt 9 Vol. 63; ISSN ACSFCL; ISSN 1744-3091
Country of Publication:
United Kingdom
Language:
English

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