TY - JOUR
T1 - Enzymatic characterization of a class II lysyl-tRNA synthetase, LysS, from Myxococcus xanthus
AU - Oka, Manami
AU - Takegawa, Kaoru
AU - Kimura, Yoshio
N1 - Funding Information:
This study was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan ( 25440087 ).
Publisher Copyright:
© 2015 Elsevier Inc. All rights reserved.
PY - 2015/6/10
Y1 - 2015/6/10
N2 - Lysyl-tRNA synthetases efficiently produce diadenosine tetraphosphate (Ap4A) from lysyl-AMP with ATP in the absence of tRNA. We characterized recombinant class II lysyl-tRNA synthetase (LysS) from Myxococcus xanthus and found that it is monomeric and requires Mn2+ for the synthesis of Ap4A. Surprisingly, Zn2+ inhibited enzyme activity in the presence of Mn2+. When incubated with ATP, Mn2+, lysine, and inorganic pyrophosphatase, LysS first produced Ap4A and ADP, then converted Ap4A to diadenosine triphosphate (Ap3A), and finally converted Ap3A to ADP, the end product of the reaction. Recombinant LysS retained Ap4A synthase activity without lysine addition. Additionally, when incubated with Ap4A (minus pyrophosphatase), LysS converted Ap4A mainly ATP and AMP, or ADP in the presence or absence of lysine, respectively. These results demonstrate that M. xanthus LysS has different enzymatic properties from class II lysyl-tRNA synthetases previously reported.
AB - Lysyl-tRNA synthetases efficiently produce diadenosine tetraphosphate (Ap4A) from lysyl-AMP with ATP in the absence of tRNA. We characterized recombinant class II lysyl-tRNA synthetase (LysS) from Myxococcus xanthus and found that it is monomeric and requires Mn2+ for the synthesis of Ap4A. Surprisingly, Zn2+ inhibited enzyme activity in the presence of Mn2+. When incubated with ATP, Mn2+, lysine, and inorganic pyrophosphatase, LysS first produced Ap4A and ADP, then converted Ap4A to diadenosine triphosphate (Ap3A), and finally converted Ap3A to ADP, the end product of the reaction. Recombinant LysS retained Ap4A synthase activity without lysine addition. Additionally, when incubated with Ap4A (minus pyrophosphatase), LysS converted Ap4A mainly ATP and AMP, or ADP in the presence or absence of lysine, respectively. These results demonstrate that M. xanthus LysS has different enzymatic properties from class II lysyl-tRNA synthetases previously reported.
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U2 - 10.1016/j.abb.2015.05.014
DO - 10.1016/j.abb.2015.05.014
M3 - Article
C2 - 26048731
AN - SCOPUS:84930959480
SN - 0003-9861
VL - 579
SP - 33
EP - 39
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
ER -