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Profiling lethal factor interacting proteins from human stomach using T7 phage display screening

  • Authors:
    • Albin Cardona‑Correa
    • Carlos Rios‑Velazquez
  • View Affiliations / Copyright

    Affiliations: Department of Biology, College of Arts and Sciences, University of Puerto Rico‑Mayagüez, Mayagüez 00681‑9000, PR, USA
    Copyright: © Cardona‑Correa et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 3797-3804
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    Published online on: March 21, 2016
       https://doi.org/10.3892/mmr.2016.5031
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Abstract

The anthrax lethal factor (LF) is a zinc dependent metalloproteinase that cleaves the majority of mitogen-activated protein kinase kinases and a member of NOD-like receptor proteins, inducing cell apoptosis. Despite efforts to fully understand the Bacillus anthracis toxin components, the gastrointestinal (GI) anthrax mechanisms have not been fully elucidated. Previous studies demonstrated gastric ulceration, and a substantial bacterial growth rate in Peyer's patches. However, the complete molecular pathways of the disease that results in tissue damage by LF proteolytic activity remains unclear. In the present study, to identify the profile of the proteins potentially involved in GI anthrax, protein‑protein interactions were investigated using human stomach T7 phage display (T7PD) cDNA libraries. T7PD is a high throughput technique that allows the expression of cloned DNA sequences as peptides on the phage surface, enabling the selection and identification of protein ligands. A wild type and mutant LF (E687A) were used to differentiate interaction sites. A total of 124 clones were identified from 194 interacting‑phages, at both the DNA and protein level, by in silico analysis. Databases revealed that the selected candidates were proteins from different families including lipase, peptidase‑A1 and cation transport families, among others. Furthermore, individual T7PD candidates were tested against LF in order to detect their specificity to the target molecule, resulting in 10 LF‑interacting peptides. With a minimum concentration of LF for interaction at 1 µg/ml, the T7PD isolated pepsin A3 pre‑protein (PAP) demonstrated affinity to both types of LF. In addition, PAP was isolated in various lengths for the same protein, exhibiting common regions following PRALINE alignment. These findings will help elucidate and improve the understanding of the molecular pathogenesis of GI anthrax, and aid in the development of potential therapeutic agents.
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Copy and paste a formatted citation
Spandidos Publications style
Cardona‑Correa A and Rios‑Velazquez C: Profiling lethal factor interacting proteins from human stomach using T7 phage display screening. Mol Med Rep 13: 3797-3804, 2016.
APA
Cardona‑Correa, A., & Rios‑Velazquez, C. (2016). Profiling lethal factor interacting proteins from human stomach using T7 phage display screening. Molecular Medicine Reports, 13, 3797-3804. https://doi.org/10.3892/mmr.2016.5031
MLA
Cardona‑Correa, A., Rios‑Velazquez, C."Profiling lethal factor interacting proteins from human stomach using T7 phage display screening". Molecular Medicine Reports 13.5 (2016): 3797-3804.
Chicago
Cardona‑Correa, A., Rios‑Velazquez, C."Profiling lethal factor interacting proteins from human stomach using T7 phage display screening". Molecular Medicine Reports 13, no. 5 (2016): 3797-3804. https://doi.org/10.3892/mmr.2016.5031
Copy and paste a formatted citation
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Spandidos Publications style
Cardona‑Correa A and Rios‑Velazquez C: Profiling lethal factor interacting proteins from human stomach using T7 phage display screening. Mol Med Rep 13: 3797-3804, 2016.
APA
Cardona‑Correa, A., & Rios‑Velazquez, C. (2016). Profiling lethal factor interacting proteins from human stomach using T7 phage display screening. Molecular Medicine Reports, 13, 3797-3804. https://doi.org/10.3892/mmr.2016.5031
MLA
Cardona‑Correa, A., Rios‑Velazquez, C."Profiling lethal factor interacting proteins from human stomach using T7 phage display screening". Molecular Medicine Reports 13.5 (2016): 3797-3804.
Chicago
Cardona‑Correa, A., Rios‑Velazquez, C."Profiling lethal factor interacting proteins from human stomach using T7 phage display screening". Molecular Medicine Reports 13, no. 5 (2016): 3797-3804. https://doi.org/10.3892/mmr.2016.5031
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