Accession Number:

AD1089845

Title:

Targeting Shiga Toxins for Treatment-resistant Bacterial Gut Infections

Descriptive Note:

Technical Report,01 Jul 2018,30 Jun 2019

Corporate Author:

University of Utah Salt Lake City United States

Personal Author(s):

• Kay,Michael S

Report Date:

2019-07-01

Pagination or Media Count:

29.0

Abstract:

The purpose of this research is to identify a protease-resistant D-peptide inhibitor of Shiga toxin for use as an orally delivered prevention or therapeutic agent to treat severe bacterial gut infections caused by Shigella or Shiga toxin-producing E. coli STEC. During this period, we successfully synthesized the Shiga toxin B-subunit using solid-phase peptide synthesis and native chemical ligation. The N-terminal half of the protein suffers from poor solubility, so we developed a new helping hand solubilizing tag to enable isolation of this peptide and its ligation to the C-terminal half. This synthetic protein was folded into pentamers and validated using LCMS, circular dichroism, size-exclusion chromatography, and analytical ultracentrifugation. To overcome problems with instability of the pentamer, we developed a cork peptide that mimics the C-terminal tail of the natural Shiga A subunit. This peptide will stabilize the B-subunit pentamer by filling its central cavity. This stabilized pentamer will be used in mirror-image phage display to identify D-peptide inhibitors.

Descriptors:

• ANTI BACTERIAL AGENTS
• drug resistance
• mass spectrometry
• wound infections
• inhibitors
• chromatography
• amines
• amino acids
• peptides
• toxins
• ESCHERICHIA COLI

Subject Categories:

• Microbiology
• Microbiology

Distribution Statement:

APPROVED FOR PUBLIC RELEASE