Part:BBa_K2688003
cpg2_tu
Executive summary : This BioBrick is a full translation unit for cpg2, an enzyme that degrades the anticancer drug methotrexate. When expressed in E. coli, it can rapidly eliminate methotrexate from the culture medium. This is proven conclusively by HPLC, and a bioassay that measures residual toxicity.
The carboxypeptidase G2 is an hydrolase that cleaves the (poly)glutamate tail off folates and analogues, leaving behind a pteroate ring system (Rowsell et al., 1997). There is broad substrate specificity, including endogenous folates, and notably the chemotherapeutic drug methotrexate (MTX). Its metabolite DAMPA has little antifolate activity in vitro, and none of the relevant clinical effects of the parent drug (B C Widemann et al., 2000). This has led to its use as both an antidote in case of MTX poisoning (Brigitte C. Widemann et al., 2010), and as a general purpose platform for novel drug delivery methods, where carboxypeptidase G2 would activate polyglutamated, soluble prodrugs in situ (Masterson et al., 2006).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 132
Illegal NgoMIV site found at 820 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 919
Functional Parameters
| ec_num | 3.4.17.11 |
| ligands | folates, folate analogues, various polyglutamated small molecules, various short peptides |
| uniprot | P06621 |
//cds/enzyme
//chassis/prokaryote/ecoli
//function/degradation
| ec_num | 3.4.17.11 |
| ligands | folates, folate analogues, various polyglutamated small molecules, various short peptides |
| uniprot | P06621 |