[Isolation and characteristics of functionally active proteolytically modified forms of tyrosyl-tRNA synthetase from bovine liver].
Functionally active proteolytic modified form of tyrosyl-tRNA-synthetase has been isolated in a homogeneous form from the bovine liver under incomplete blocking of endogenous proteolysis. The isolation scheme is described. From the data of gel electrophoresis under denaturing conditions the molecular weight of this form is 39 +/- 1.5 kDa and from the data of gel filtration under native conditions -84 kDa. Thus, this form as well as the native enzyme is a dimer of the alpha 2-type. As compared to the native enzyme (Mm 2 x 59 kDa) a proteolytically modified form has a fragment of the polypeptide chain about 20 kDa long split out (this fragment is not essential for catalytic activity). The values of catalytic characteristics of the modified form in tRNA(Tyr) aminoacylation reaction (Km = 1.19 microM and kcat = 2.99 min-1) are close to those obtained for the main form of the enzyme (0.69 microM and 2.97 min-1, respectively). Amino acid composition of the low-molecular form of tyrosyl-tRNA-synthetase has been determined. It was found that the fragment split out in limited proteolysis was characterized by very high content of positively charged lysine residues (46 residues). A proteolytically modified form of tyrosyl-tRNA-synthetase possesses, like the main form, the affinity to high-molecular rRNA but it is eluted from the column filled with rRNA-sepharose at lower salt concentration (50 mM KCl) as compared to the main form of the enzyme (100 mM KCl).
Gnatenko
D V
DV
Korneliuk
A I
AI
Kurochkin
I V
IV
Ribkinska
T A
TA
Matsuka
G Kh
GKh
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002384:Catalysis; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008099:Liver / Q000201:enzymology*; D011487:Protein Conformation; D014448:Tyrosine-tRNA Ligase / Q000302:isolation & purification* / Q000378:metabolism
D000596:Amino Acids; D014448:Tyrosine-tRNA Ligase
Vydelenie i kharkteristkia funktsional'no aktivnoĭ proteoliticheski modifitsirovannoĭ formy tirozil-tRNK-sintetazy iz pecheni byka.