Part:BBa_K5057013
Sushi S1 antimicrobial peptide with periplasmic signal peptide pelB
Sushi S1 is an antimicrobial cationic peptide composed of 34 amino acids, derived from the lipopolysaccharide (LPS)-binding region of Factor C found in horseshoe crabs. It targets bacterial membranes through four successive steps in the bactericidal process: 1) Binding, primarily mediated by charged residues in the peptide; 2) Peptide association; 3) Membrane disruption, during which lipopolysaccharides remain intact; and 4) Lysis, resulting from the leakage of cytosolic contents through large membrane defects.
In our experiments, we investigated the effect of intracellular expression of pelB-Sushi S1 (Biobricks: BBa_J32015, BBa_K5057004 ) encoded on pET22b(+) backbone on the viability of bacterial cultures, as well as the impact of the purified peptide on the bacterial growth.
Usage and Biology
Antimicrobial peptides (AMPs) are a diverse class of small, naturally occurring peptides playing a crucial role in the innate immune response of various organisms. These peptides consist of 10 to 60 amino acids and are generally characterized by their net positive charge and the ability to disrupt microbial membranes, thereby exhibiting potent activity against a wide range of pathogens, including bacteria, fungi, viruses and parasites. Most AMPs target bacterial membranes by creating pores or disrupting the whole membranes in the detergent-like manner [1]. These modes of action rely on both the cationic properties of the AMP itself, and negatively charged bacterial membranes. The differences in lipid composition between the host and pathogen membranes enable the AMPs to achieve high specificity [2].
Ongoing research has led to the discovery of further categories of AMPs exhibiting inhibitory or disruptive effects on protein and DNA synthesis, cell division and biofilm formation. These peptides rely on various mechanisms involving enzyme inactivation, signaling disruption or induction of degradation processes [3, 4, 5, 6].
Due to their extraoridinary characteristics, AMPs constitute a promising research field for the development of new therapeutics to combat antibiotic resistance. Many attempts have been made to create synthetic AMPs de novo, mimicking the design of already existing peptides [7, 8, 9]. However, designing AMPs comes with several obstacles effectively preventing a wider use of AMPs in the medicine. As stated by Li et al.[2] AMPs often have a hemolytic effect on eukaryotic cells, they lack stability due to limited pH tolerance and proteolyse susceptibility and experience reduced activity in the presence of iron and different serums. A further limitation is also high costs of AMP production typically by chemical synthesis [10, 2].
Sushi S1 derived from factor C, a protein involved in the coagulation cascade of the horseshoe crab, has shown rapid bactericidal effect and high affinity for LPS present on the outer membrane of various Gram-negative bacteria including E. coli and Pseudomonas aeruginosa. Sushi S1 may neutralize LPS biotoxicity and mitigate the severe effects of septic shock - a condition that arises from bacterial infections and consecutive antibiotics treatment and is characterized by critical health complications, particularly in vulnerable populations. Furthermore, Sushi S1 shows reportedly low cytotoxic activity against mammalian erythrocytes and remains active in a physiologically relatively broad pH range (pH 6-8) and osmolarity ( 50 to 300 mM) [11].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 54
- 1000COMPATIBLE WITH RFC[1000]
References
[1] Huan Y, Kong Q, Mou H, Yi H. Antimicrobial Peptides: Classification, Design, Application and Research Progress in Multiple Fields. Frontiers in Microbiology [Internet]. 2020 Oct 16;11.
[2] Li J, Koh JJ, Liu S, Lakshminarayanan R, Verma CS, Beuerman RW. Membrane Active Antimicrobial Peptides: Translating Mechanistic Insights to Design. Frontiers in Neuroscience. 2017 Feb 14;11.
[3] Mardirossian M, Pérébaskine N, Benincasa M, Stefano Gambato, Hofmann S, Huter P, et al. The Dolphin Proline-Rich Antimicrobial Peptide Tur1A Inhibits Protein Synthesis by Targeting the Bacterial Ribosome. 2018 May 17;25(5):530-539.e7.
[4] He SW, Zhang J, Li NQ, Zhou S, Yue B, Zhang M. A TFPI-1 peptide that induces degradation of bacterial nucleic acids, and inhibits bacterial and viral infection in half-smooth tongue sole, Cynoglossus semilaevis. Fish & Shellfish Immunology [Internet]. 2017 Jan 1;60:466–73.
[5] Lutkenhaus J. Regulation of cell division in E. coli. Trends in Genetics. 1990;6:22–5.
[6] Li L, Sun J, Xia S, Tian X, Cheserek MJ, Le G. Mechanism of antifungal activity of antimicrobial peptide APP, a cell-penetrating peptide derivative, against Candida albicans: intracellular DNA binding and cell cycle arrest. Applied Microbiology and Biotechnology. 2016 Jan 8;100(7):3245–53.
[7] Goormaghtigh E, Meutter J, Szoka F, Cabiaux V, Parente RA, Ruysschaert JM. Secondary structure and orientation of the amphipathic peptide GALA in lipid structures. An infrared-spectroscopic approach. European Journal of Biochemistry. 1991 Jan;195(2):421–9.
[8] Fjell CD, Hiss JA, Hancock REW, Schneider G. Designing antimicrobial peptides: form follows function. Nature Reviews Drug Discovery. 2011 Dec 16;11(1):37–51.
[9] Jenisha G, Hart RJ, Soldano A, Chen CH, Guha S, Hoffmann JP, et al. Optimization of Host Cell-Compatible, Antimicrobial Peptides Effective against Biofilms and Clinical Isolates of Drug-Resistant Bacteria. ACS Infectious Diseases [Internet]. 2023 Mar 24;(4):952–65.
[10] Jaradat DMM. Thirteen decades of peptide synthesis: key developments in solid phase peptide synthesis and amide bond formation utilized in peptide ligation. Amino Acids. 2017 Nov 28;50(1):39–68.
[11] Yau YH, Ho B, Tan NS, Ng ML, Ding JL. High therapeutic index of factor C Sushi peptides: potent antimicrobials against Pseudomonas aeruginosa. Antimicrob Agents Chemother [Internet]. 2001;45(10):2820–5. Available from: http://dx.doi.org/10.1128/AAC.45.10.2820-2825.2001
| None |