PDBsum entry 4jhz

Hydrolase/hydrolase inhibitor

PDB id: 4jhz

Contents

Protein chains

596 a.a.

Ligands

1KV ×2

Waters ×124

PDB id:

4jhz

Name:

Hydrolase/hydrolase inhibitor

Title:

Structure of e. Coli beta-glucuronidase bound with a novel, potent inhibitor 2-[4-(1,3-benzodioxol-5-ylmethyl)piperazin-1-yl]-n-[(1s,2s, 5s)-2,5-dimethoxycyclohexyl]acetamide

Structure:

Beta-glucuronidase. Chain: a, b. Synonym: gus, beta-d-glucuronoside glucuronosohydrolase. Engineered: yes

Source:

Escherichia coli. Organism_taxid: 83333. Strain: k12. Gene: b1617, gura, gusa, jw1609, uida. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

2.83Å R-factor: 0.198 R-free: 0.247

Authors:

A.B.Roberts,B.D.Wallace,M.R.Redinbo

Key ref:

A.B.Roberts et al. (2013). Molecular insights into microbial β-glucuronidase inhibition to abrogate CPT-11 toxicity. Mol Pharmacol, 84, 208-217. PubMed id: 23690068 DOI: 10.1124/mol.113.085852

Date:

05-Mar-13 Release date: 28-Aug-13

Protein chains

P05804  (BGLR_ECOLI) -  Beta-glucuronidase from Escherichia coli (strain K12)

Seq: 603 a.a.

Struc: 596 a.a.

Enzyme reactions

• Enzyme class: E.C.3.2.1.31 - beta-glucuronidase.
• Reaction: a beta-D-glucuronoside + H2O = D-glucuronate + an alcohol

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1124/mol.113.085852

Mol Pharmacol 84:208-217 (2013)

PubMed id: 23690068

Molecular insights into microbial β-glucuronidase inhibition to abrogate CPT-11 toxicity.

A.B.Roberts, B.D.Wallace, M.K.Venkatesh, S.Mani, M.R.Redinbo.

ABSTRACT

Bacterial β-glucuronidases expressed by the symbiotic intestinal microbiota appear to play important roles in drug-induced epithelial cell toxicity in the gastrointestinal (GI) tract. For the anticancer drug CPT-11 (irinotecan) and the nonsteroidal anti-inflammatory drug diclofenac, it has been shown that removal of the glucuronide moieties from drug metabolites by bacterial β-glucuronidases in the GI lumen can significantly damage the intestinal epithelium. Furthermore, selective disruption of bacterial β-glucuronidases by small molecule inhibitors alleviates these side effects, which, for CPT-11 {7-ethyl-10-[4-(1-piperidino)-1-piperidino]}, can be dose limiting. Here we characterize novel microbial β-glucuronidase inhibitors that inhibit Escherichia coli β-glucuronidase in vitro with Ki values between 180 nM and 2 μM, and disrupt the enzyme in E. coli cells, with EC50 values as low as 300 nM. All compounds are selective for E. coli β-glucuronidase without inhibiting purified mammalian β-glucuronidase, and they do not impact the survival of either bacterial or mammalian cells. The 2.8 Å resolution crystal structure of one inhibitor bound to E. coli β-glucuronidase demonstrates that it contacts and orders only a portion of the "bacterial loop" present in microbial, but not mammalian, β-glucuronidases. The most potent compound examined in this group was found to protect mice against CPT-11-induced diarrhea. Taken together, these data advance our understanding of the chemical and structural basis of selective microbial β-glucuronidase inhibition, which may improve human drug efficacy and toxicity.