Difference between revisions of "Part:BBa K2688003"

Revision as of 10:51, 6 October 2018

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 132
Illegal NgoMIV site found at 820
1000
INCOMPATIBLE 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

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2688003 short</partinfo>
-This composite part is a full translation unti for the Carboxypeptidase G2 (cpg2) without periplasmic localization signal.
+'''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).
-This enzyme cleaves folates into pteroates and glutamate(s). It has broad specificity, acting on folic acid, reduced folate but also folate analogs like methotrexate.