ライフサイエンスコーパス: Asp-tRNA

1 he conversion by an amidotransferase of [14C]Asp-tRNA to [14C]Asn-tRNA.

2 thway involves the formation of mis-acylated Asp-tRNA(Asn) or Glu-tRNA(Gln), and the subsequent amida

3 on the misacylated tRNAs Glu-tRNA (Gln) and Asp-tRNA (Asn) as intermediates.

4 version of the misacylated Glu-tRNA(Gln) and Asp-tRNA(Asn) species catalyzed by the GatCAB amidotrans

5 tRNA(Pro), Ser-tRNA(Thr), Glu-tRNA(Gln), and Asp-tRNA(Asn).

6 hat Thermus thermophilus EF-Tu does not bind Asp-tRNA (Asn) and predicted a similar discriminatory re

7 discriminating (ND-AspRS) and generates both Asp-tRNA(Asp) and the noncanonical, misacylated Asp-tRNA

8 ting that Hp Asp/Glu-Adt can utilize E. coli Asp-tRNA(Asn) as a substrate.

9 In contrast, Asp-tRNA formed by the Pyrococcus or Ferroplasma enzymes

10 also enhances the capacity of AdT to convert Asp-tRNA(Asn) into Asn-tRNA(Asn) by approximately 35-fol

11 nt C. trachomatis amidotransferase converted Asp-tRNA(Asn) and Glu-tRNA(Gln) into Asn-tRNA and Gln-tR

12 equently repaired by the glutamine-dependent Asp-tRNA(Asn)/Glu-tRNA(Gln) amidotransferase (Asp/Glu-Ad

13 h the same DNA library and then screened for Asp-tRNA(Asp) formation in vivo by growth at the non-per

14 and then subjected to in vivo selection for Asp-tRNA(Asn) formation by growth in minimal medium.

15 dakaraensis AspRS gained the ability to form Asp-tRNA(Asn) in vitro when the W26H or K85P changes wer

16 rans AspRS2 enzymes still capable of forming Asp-tRNA(Asp) but unable to recognize tRNA(Asn).

17 spRS), an enzyme that in addition to forming Asp-tRNA(Asp) also misacylates tRNA(Asn).

18 ating AspRS (D-AspRS) specifically generates Asp-tRNA(Asp) and usually coexists with asparaginyl-tRNA

19 Human Asp-tRNA synthetase contains a short 32-residue amino-te

20 ing the prediction that Glu-tRNA (Gln), like Asp-tRNA (Asn), will not form a complex with EF-Tu.

21 -tRNA(Asp) and the noncanonical, misacylated Asp-tRNA(Asn); this misacylated tRNA is subsequently rep

22 a two-step pathway that requires misacylated Asp-tRNA(Asn) as an intermediate.

23 or Gln-tRNA, by transamidation of mischarged Asp-tRNA(Asn) or Glu-tRNA(Gln) catalyzed by a heterotrim

24 nsferase complex (Gat CAB) genes that modify Asp-tRNA(Asn) into Asn-tRNA(Asn) and Glu-tRNA(Gln) into

25 on in most prokaryotes requires amidation of Asp-tRNA or Glu-tRNA by amidotransferases that couple an

26 While large amounts of Asp-tRNA(Asn) are detrimental to E. coli, smaller amount

27 methanogen synthetase was the conversion of Asp-tRNA formed by M. thermautotrophicus AspRS to Asn-tR

28 This complex enables direct delivery of Asp-tRNA(Asn) from the non-discriminating aspartyl-tRNA

29 th which to measure the in vivo formation of Asp-tRNA(Asn) and its acceptance by elongation factor EF

30 e in the genome of AsnRS and the presence of Asp-tRNA(Asn) amidotransferase, employed by the transami

31 The presence of Asp-tRNA(Asn) and ATP enhances the glutaminase activity

32 phoresis confirmed the in vitro synthesis of Asp-tRNA(Asn).

33 ged by the ratio of the k(cat)K(m) values of Asp-tRNA(Asp) vs. Asp-tRNA(Asn) formation.

34 e discriminating enzyme (D-AspRS) forms only Asp-tRNA(Asp), whereas the nondiscriminating enzyme (ND-

35 e discriminating enzyme (D-AspRS) forms only Asp-tRNA(Asp), while the nondiscriminating one (ND-AspRS

36 contrast, a discriminating AspRS forms only Asp-tRNA(Asp).

37 at does not utilize either Glu-tRNA (Gln) or Asp-tRNA (Asn), and the second from Helicobacter pylori,

38 he mischarged tRNA species, Glu-tRNA(Gln) or Asp-tRNA(Asn).

39 Most prokaryotes require Asp-tRNA(Asn) for the synthesis of Asn-tRNA(Asn).

40 scriminating one (ND-AspRS) also synthesizes Asp-tRNA(Asn), a required intermediate in protein synthe

41 iminating enzyme (ND-AspRS) also synthesizes Asp-tRNA(Asn), which is a required intermediate for prot

42 Proof that Asp-tRNA(Asn) was generated by the methanogen synthetase

43 Northern blot analysis of the Asp-tRNA separated by acidurea gel electrophoresis confi

44 s AspRS to Asn-tRNA by M. thermautotrophicus Asp-tRNA(Asn) amidotransferase.

45 hat while the enzyme is able to transamidate Asp-tRNA(Asn) (k(cat)/K(M) of 125 s(-1)/mM) it is unable

46 zation reveals that the enzyme transamidates Asp-tRNA(Asn) and Glu-tRNA(Gln) with similar efficiency

47 f the k(cat)K(m) values of Asp-tRNA(Asp) vs. Asp-tRNA(Asn) formation.