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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli | |
Zn2+ | required for diadenosine tetraphosphate (Ap4A) formation | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-lysine + tRNALys | Escherichia coli | - |
AMP + diphosphate + L-lysyl-tRNALys | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A8N5 | - |
- |
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 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
LysU | - |
Escherichia coli |
lysyl tRNA synthetase | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli |
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 |