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Literature summary for 6.1.1.6 extracted from

  • Wright, M.; Azhar, M.A.; Kamal, A.; Miller, A.D.
    Syntheses of stable, synthetic diadenosine polyphosphate analogues using recombinant histidine-tagged lysyl tRNA synthetase (LysU) (2014), Bioorg. Med. Chem. Lett., 24, 2346-2352 .
    View publication on PubMed
Search for more literature for 6.1.1.6

Application

Application Comment Organism
drug development LysU is also useful as a tool for highly controlled phosphate-phosphate bond formation between nucleotides, avoiding the need for complex protecting group chemistries. Resulting high yielding tandem LysU-based biosynthetic-synthetic/synthetic-biosynthetic strategies emerge for the preparation of varieties of ApnA analogues directly from inexpensive natural nucleotides and nucleosides. Analogues so formed make a useful small library with which to probe ApnA activities in vitro and in vivo leading to the discovery of potentially potent biopharmaceuticals active against chronic pain and other chronic, high-burden disease states Escherichia coli
synthesis recombinant Escherichia coli lysyl-tRNA synthase (LysU) has been previously utilised in the production of stabile, synthetic diadenosine polyphosphate (ApnA) analogues. LysU is also useful as a tool for highly controlled phosphate-phosphate bond formation between nucleotides, avoiding the need for complex protecting group chemistries. Resulting high yielding tandem LysU-based biosynthetic-synthetic/synthetic-biosynthetic strategies emerge for the preparation of varieties of ApnA analogues directly from inexpensive natural nucleotides and nucleosides. Analogues so formed make a useful small library with which to probe ApnA activities in vitro and in vivo leading to the discovery of potentially potent biopharmaceuticals active against chronic pain and other chronic, high-burden disease states Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Escherichia coli
Zn2+ required for diadenosine tetraphosphate (Ap4A) formation Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-lysine + tRNALys Escherichia coli
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 AMP + diphosphate + L-lysyl-tRNALys
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 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A8N5
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-lysine + tRNALys
-
 Escherichia coli AMP + diphosphate + L-lysyl-tRNALys
-
 ?
additional information LysU-based preparation of potentially important ApnA analogues, overview. Dimeric LysU has dual diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) and diadenosine-5',5'''-P1,P3-triphosphate (Ap3A) synthase activities. Syntheses of both take place through the formation of a lysyl-adenylate 1 intermediate from ATP and L-lysine. Thereafter, the terminal phosphate of a second nucleotide substrate combines with the enzyme-bound lysyl-adenylate, thereby liberating free L-lysine and generating either Ap4A or Ap3A depending upon the identity of the second nucleotide substrate. The first step involving lysyladenylate intermediate formation is highly specific but reversible. Therefore inorganic diphosphatase-mediated controlled hydrolysis of diphosphate is required in order to prevent the back-reaction taking place, and thereby essentially rendering this first step committed. Fortunately, the second product formation step is highly promiscuous and a wide variety of nucleotide di-, tri-, and tetraphosphates are acceptable as second nucleotide substrates. This promiscuity also extends to inorganic phosphate and to tripolyphosphate. Surface mechanism of LysU catalyzed Ap4A and Ap3A synthase activities, reaction scheme and mechanism, overview. Bulkier putative diphosphate analogue substrates preclude molecular recognition and binding by LysU, hence preventing their use as bona fide LysU substrates able to couple to the ATP derived lysyl adenylate 1 intermediate with LysU assistance. Synthesis of analogues beta,gamma-methylene-P1,P4-bis(5'-adenosyl) tetraphosphate, beta,gamma-imido-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)2-beta,gamma-methylene-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)2-beta,gamma-imido-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)-beta,gamma-methylene-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)-beta,gamma-imido-P1,P4-bis(5'-adenosyl) tetraphosphate, alpha,beta-methylene 5'-P1,P3-bis(5'-adenosyl) triphosphate, alpha,beta-methylene-guanosine 5'-P1-triphospho-P3-5''-adenosine, beta,gamma-methylene-P1,P5-bis(5'-adenosyl) pentaphosphate, beta,gamma-imido-adenosine 5'-P1-pentaphospho-P5-5''-uridine, beta,gamma-methylene-adenosine 5'-P1-tretraphospho-P4-5''-guanosine, beta,gamma-imido-adenosine 5'-P1-tretraphospho-P4-5''-guanosine, and beta,gamma-delta,epsilon-dimethylene-P1,P6-bis(5'-adenosyl) hexaphosphate Escherichia coli ?
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 ?

Synonyms

Synonyms Comment Organism
LysU
-
 Escherichia coli
lysyl tRNA synthetase
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Escherichia coli

General Information

General Information Comment Organism
additional information LysU is useful as a tool for highly controlled phosphate-phosphate bond formation between nucleotides, avoiding the need for complex protecting group chemistries. Resulting high yielding tandem LysU-based biosynthetic-synthetic/synthetic-biosynthetic strategies emerge for the preparation of varieties of ApnA analogues directly from inexpensive natural nucleotides and nucleosides. Analogues so formed make a useful small library with which to probe ApnA activities in vitro and in vivo leading to the discovery of potentially potent biopharmaceuticals active against chronic pain and other chronic, high-burden disease states Escherichia coli