Purification, crystallization and preliminary X-ray characterization of a human mitochondrial phenylalanyl-tRNA synthetase.
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 alpha-subunit of PheRS and the B8 domain of its beta-subunit. Together, the alpha-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 A resolution and belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 55, b = 90, c = 96 A.
Levin
Inna
I
Department of Structural Biology, Weizmann Institute of Science, 76100 Rehovot, Israel.
Kessler
Naama
N
Moor
Nina
N
Klipcan
Liron
L
Koc
Emine
E
Templeton
Paul
P
Spremulli
Linda
L
Safro
Mark
M
D000255:Adenosine Triphosphate / Q000378:metabolism; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008928:Mitochondria / Q000201:enzymology*; D010649:Phenylalanine / Q000378:metabolism; D010652:Phenylalanine-tRNA Ligase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D014961:X-Ray Diffraction
D010649:Phenylalanine; D000255:Adenosine Triphosphate; D010652:Phenylalanine-tRNA Ligase