alexa Fluorescence High-throughput Screening For Inhibitors Of TonB Action | 71892
ISSN: 2332-0877

Journal of Infectious Diseases & Therapy
Open Access

OMICS International organises 3000+ Global Conferenceseries Events every year across USA, Europe & Asia with support from 1000 more scientific Societies and Publishes 700+ Open Access Journals which contains over 50000 eminent personalities, reputed scientists as editorial board members.

Open Access Journals gaining more Readers and Citations
700 Journals and 15,000,000 Readers Each Journal is getting 25,000+ Readers

This Readership is 10 times more when compared to other Subscription Journals (Source: Google Analytics)

3rd Annual Congress on Infectious Diseases

Phillip E Klebba, Olivia S Eliasson, Dallas R Hyder, Noah J Long, Aritri Majumdar, Somnath, Chakravorty, Peter McDonald, Anuradha Roy, Salete M Newton and Brittany L Nairn
Kansas State University, USA
Posters & Accepted Abstracts: J Infect Dis Ther
DOI: 10.4172/2332-0877-C1-027
Abstract
Gram (-) bacteria acquire ferric siderophores through TonB-dependent transporters (TBDT) in their outer membrane. By fluorescence spectroscopic high-throughput screening (FLHTS) we identified inhibitors of TonB-dependent ferric enterobactin (FeEnt) uptake through E. coli FepA (EcoFepA). Among 165 inhibitors found in a primary screen of 17,441 compounds, we evaluated 20 candidates in secondary tests of TonB activity, including ferric siderophore uptake and colicin killing. 6 of the 20 primary hits inhibited TonB action in all the tests. Further analysis of the inhibitors in [59Fe] Ent and [14C]- lactose accumulation experiments suggested several as proton ionophores, but two chemicals, ebselen and ST0082990, did not behave like proton ionophores and may inhibit TonB-ExbBD. The success of FLHTS against E. coli led us to adapt it to the ESKAPE pathogen Acinetobacter baumannii. We identified its FepA ortholog (AbaFepA), confirmed its involvement in FeEnt uptake by deleting the structural gene, cloned AbafepA, genetically engineered 8 Cys substitutions in its surface loops, modified them with fluorescein and made fluorescence spectroscopic observations of FeEnt uptake in A. baumannii. Among the Cys substitutions in AbaFepA, several (S279C, T562C, S665C) were well labeled by fluorescein and suitable for measurements of FeEnt transport. As in E. coli, the test monitored TonB-dependent FeEnt uptake by AbaFepA. In micro titer format FLHTS with A. baumannii produced Z’ factors from 0.6-0.8. Overall these experiments both identified agents that block TonB action, and revealed the potential of FLHTS for larger screens of bigger libraries to find novel antimicrobial compounds against Gram (-) bacteria, including the CRE/ESKAPE pathogens.
image PDF   |   image HTML
 

Relevant Topics

Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2018-19
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

Agri & Aquaculture Journals

Dr. Krish

[email protected]

1-702-714-7001Extn: 9040

Biochemistry Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Business & Management Journals

Ronald

[email protected]

1-702-714-7001Extn: 9042

Chemistry Journals

Gabriel Shaw

[email protected]

1-702-714-7001Extn: 9040

Clinical Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Engineering Journals

James Franklin

[email protected]

1-702-714-7001Extn: 9042

Food & Nutrition Journals

Katie Wilson

[email protected]

1-702-714-7001Extn: 9042

General Science

Andrea Jason

[email protected]

1-702-714-7001Extn: 9043

Genetics & Molecular Biology Journals

Anna Melissa

[email protected]

1-702-714-7001Extn: 9006

Immunology & Microbiology Journals

David Gorantl

[email protected]

1-702-714-7001Extn: 9014

Materials Science Journals

Rachle Green

[email protected]

1-702-714-7001Extn: 9039

Nursing & Health Care Journals

Stephanie Skinner

[email protected]

1-702-714-7001Extn: 9039

Medical Journals

Nimmi Anna

[email protected]

1-702-714-7001Extn: 9038

Neuroscience & Psychology Journals

Nathan T

[email protected]

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

Ann Jose

[email protected]

1-702-714-7001Extn: 9007

Social & Political Science Journals

Steve Harry

[email protected]

1-702-714-7001Extn: 9042

 
© 2008- 2018 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version