dc.contributor.author |
Williams, Elsie M |
|
dc.contributor.author |
Rich, Michelle H |
|
dc.contributor.author |
Mowday, Alexandra M |
|
dc.contributor.author |
Ashoorzadeh, Amir |
|
dc.contributor.author |
Copp, Janine N |
|
dc.contributor.author |
Guise, Christopher P |
|
dc.contributor.author |
Anderson, Robert F |
|
dc.contributor.author |
Flanagan, Jack U |
|
dc.contributor.author |
Smaill, Jeff B |
|
dc.contributor.author |
Patterson, Adam V |
|
dc.contributor.author |
Ackerley, David F |
|
dc.date.accessioned |
2020-12-09T02:42:02Z |
|
dc.date.available |
2020-12-09T02:42:02Z |
|
dc.date.issued |
2019-9 |
|
dc.identifier.issn |
0006-2960 |
|
dc.identifier.uri |
http://hdl.handle.net/2292/54001 |
|
dc.description.abstract |
Gene-directed enzyme prodrug therapy (GDEPT) uses tumor-tropic vectors to deliver prodrug-converting enzymes such as nitroreductases specifically to the tumor environment. The nitroreductase NfsB from Escherichia coli (NfsB_Ec) has been a particular focal point for GDEPT and over the past 25 years has been the subject of several engineering studies seeking to improve catalysis of prodrug substrates. To facilitate clinical development, there is also a need to enable effective non-invasive imaging capabilities. SN33623, a 5-nitroimidazole analogue of 2-nitroimidazole hypoxia probe EF5, has potential for PET imaging exogenously delivered nitroreductases without generating confounding background due to tumor hypoxia. However, we show here that SN33623 is a poor substrate for NfsB_Ec. To address this, we used assay-guided sequence and structure analysis to identify two conserved residues that block SN33623 activation in NfsB_Ec and close homologues. Introduction of the rational substitutions F70A and F108Y into NfsB_Ec conferred high levels of SN33623 activity and enabled specific labeling of E. coli expressing the engineered enzyme. Serendipitously, the F70A and F108Y substitutions also substantially improved activity with the anticancer prodrug CB1954 and the 5-nitroimidazole antibiotic prodrug metronidazole, which is a potential biosafety agent for targeted ablation of nitroreductase-expressing vectors. |
|
dc.format.medium |
Print-Electronic |
|
dc.language |
eng |
|
dc.publisher |
American Chemical Society (ACS) |
|
dc.relation.ispartofseries |
Biochemistry |
|
dc.rights |
Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. |
|
dc.rights.uri |
https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm |
|
dc.subject |
HCT116 Cells |
|
dc.subject |
Humans |
|
dc.subject |
Escherichia coli |
|
dc.subject |
Neoplasms |
|
dc.subject |
Hydrocarbons, Fluorinated |
|
dc.subject |
Etanidazole |
|
dc.subject |
Imidazoles |
|
dc.subject |
Nitroreductases |
|
dc.subject |
Escherichia coli Proteins |
|
dc.subject |
Antineoplastic Agents |
|
dc.subject |
Prodrugs |
|
dc.subject |
Positron-Emission Tomography |
|
dc.subject |
Drug Monitoring |
|
dc.subject |
Protein Engineering |
|
dc.subject |
Biosensing Techniques |
|
dc.subject |
Cell Hypoxia |
|
dc.subject |
Enzyme Activation |
|
dc.subject |
Molecular Imaging |
|
dc.subject |
Genetic Therapy |
|
dc.subject |
Science & Technology |
|
dc.subject |
Life Sciences & Biomedicine |
|
dc.subject |
Biochemistry & Molecular Biology |
|
dc.subject |
GENE-THERAPY |
|
dc.subject |
CELL SENSITIZATION |
|
dc.subject |
POSITIVE-SELECTION |
|
dc.subject |
PHASE-I |
|
dc.subject |
NITROREDUCTASE |
|
dc.subject |
ZEBRAFISH |
|
dc.subject |
ABLATION |
|
dc.subject |
CANCER |
|
dc.subject |
EXPRESSION |
|
dc.subject |
REDUCTASE |
|
dc.subject |
0304 Medicinal and Biomolecular Chemistry |
|
dc.subject |
0601 Biochemistry and Cell Biology |
|
dc.subject |
1101 Medical Biochemistry and Metabolomics |
|
dc.title |
Engineering Escherichia coli NfsB To Activate a Hypoxia-Resistant Analogue of the PET Probe EF5 To Enable Non-Invasive Imaging during Enzyme Prodrug Therapy. |
|
dc.type |
Journal Article |
|
dc.identifier.doi |
10.1021/acs.biochem.9b00376 |
|
pubs.issue |
35 |
|
pubs.begin-page |
3700 |
|
pubs.volume |
58 |
|
dc.date.updated |
2020-11-16T18:48:50Z |
|
dc.rights.holder |
Copyright: The author |
en |
pubs.author-url |
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000484643700009&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=6e41486220adb198d0efde5a3b153e7d |
|
pubs.end-page |
3710 |
|
pubs.publication-status |
Published |
|
dc.rights.accessrights |
http://purl.org/eprint/accessRights/RestrictedAccess |
en |
pubs.subtype |
Research Support, Non-U.S. Gov't |
|
pubs.subtype |
Journal Article |
|
pubs.elements-id |
781871 |
|
dc.identifier.eissn |
1520-4995 |
|
pubs.online-publication-date |
2019-8-12 |
|