Part:BBa_K4075002
E10 Acidic Model pro-peptide
Synthetic Acidic Model Pro-domain
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Profile
Name: Acidic model pro-peptide
Base Pairs: 30 bp
Origin: Designed
Properties: Potential Acidic Model pro-peptide for Antimicrobial Peptides
Design considerations: Codon optimization for E. coli
Source: [1]
Usage and Biology
Many antimicrobial peptides (AMPs) are synthesized, delivered or stored in an inactive form, known as AMPs precursors (proAMPs), that require proteolytic cleavage to become active (i.e. cathelicidins, defensins, etc). Therefore, gene expression is not the main regulation mechanism and the abundance of appropriate proteases to activate it became a potential strategy to regulate it. These precursor forms often have a tripartite structure as peptide signal, acidic pro-peptide (or pro-domain) and the antimicrobial itself [1,2]. The acidic pro-peptide has charge densities that neutralize the positive charge of AMPs, the electrostatic neutralization turns the AMPs less toxic in an inactive form, which molecularly is still unclear [1].
This mechanism has been exploited for biotechnological applications as efficiently peptide recombinant production, reducing the toxicity to the host bacteria, or the pro-drug strategy, whereas can activate the AMP upon the presence of a specific pathogen [1].
Júnior et al. (2018) designed a synthetic deca acidic model pro-peptide based on glutamic acids (Glu x 10) to modulate the conformational change of crotalicidin, the results suggest that attaching this part leads to complete inhibition of antimicrobial activity, but it will be necessary to probe the real inactivation mechanism through of various antimicrobial peptides. In addition, the Dermaseptin pro-peptides are highly conserved, which allows the application of this pro-peptide in many Dermaseptin [3].
Therefore, the UNILA_LatAm 2021 team selected pro-peptides to act as an inactivator for leishmanicidal candidates such as CAM-W (BBa_K4075005), DRS-N1 (BBa_K4075004), DRS-S1 (BBa_K4075006) e DRS-H3 (BBa_K4075007). The composite structure contains a trypsin cleavage site between pro-peptide and AMP-activated form, this site allows the proAMPs to be split after trypsin digestion and the AMP-activated form restores the antimicrobial capacity.
| Pro-peptide | AMP | Final Composite |
|---|---|---|
| DRS-H3 pro-peptide | DRS-N1 | BBa_K4075008 |
| E10 pro-peptide | CAM-W | BBa_K4075009 |
| DRS-S1 pro-peptide | DRS-S1 | BBa_K4075010 |
| DRS-H3 pro-peptide | DRS-H3 | BBa_K4075011 |
| E10 pro-peptide | DRS-N1 | BBa_K4075012 |
[1] Júnior, N.G.O. et al. (2018). “An acidic model pro-peptide affects the secondary structure, membrane, interactions and antimicrobial activity of a crotalicidin fragment.” Nature: Scientific Reports. vol. 8:11127.
[2] Mahlapuu, M. et al. (2016). “Antimicrobial Peptides: An Emerging Category of Therapeutic Agents.” Frontiers in Cellular and Infection Microbiology. 6:194.
[3] Brand, G. D. et al. (2006) “Novel dermaseptins from Phyllomedusa hypochondrialis (Amphibia).” Biochemical and Biophysical Research Communications 347(3): 739-746.
| None |