[Express method of isolation of mammalian phenylalanine-tRNA-synthetase and preparation of monoclonal antibodies against this enzyme].
A rapid and efficient procedure for isolating homogeneous beef liver phenylalanyl-tRNA synthetase (EC.6.1.1) was developed that enables to purify the enzyme 5000 fold and to achieve the activity of 8 e.a.u. per mg of protein. The molecular mass of the native enzyme was estimated to be 260 kDa, for alpha subunit - 59 kDa, and for beta - 72 kDa. Two cellular clones were derived by means of hybridization of immunised splenocytes with myeloma cells. They secrete monoclonal antibodies, designated P6 and P1 2, that bind to human placental and bovine liver phenylalanyl-tRNA synthetases but not to the same enzymes from E. coli and T. thermophilus. P6 and P1 2 antibodies do not affect the aminoacylation capacity of human or bovine phenylalanyl-tRNA synthetases. By immunoblotting, it was shown that P6 antibodies recognize the alpha subunit of the enzyme.
O A
OA
Vartanian
A V
AV
Mashanov-Golikov
A D
AD
Vol'fson
O D
OD
Zakharova
O I
OI
Lavrik
V V
VV
Filonenko
S F
SF
Beresten'
D000604:Amino Acyl-tRNA Synthetases / Q000302:isolation & purification*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002417:Cattle; D004926:Escherichia coli / Q000201:enzymology; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D010652:Phenylalanine-tRNA Ligase / Q000276:immunology / Q000302:isolation & purification*; D013045:Species Specificity; D013824:Thermus / Q000201:enzymology
D000911:Antibodies, Monoclonal; D000604:Amino Acyl-tRNA Synthetases; D010652:Phenylalanine-tRNA Ligase
Bystryĭ metod vydeleniia fenilalanil-tRNK-sintetazy iz mlekopitaiushchikh i poluchenie monoklonal'nykh antitel k étomu fermentu.