Modification of the C terminus of cecropin is essential for broad-spectrum antimicrobial activity

Abstract

Cecropin A is a naturally occurring peptide with bactericidal activity against gram-negative and grampositive bacteria. Production of large quantities of bactericidal peptides that are similar in structure and activity to cecropin A has been achieved by combining recombinant DNA techniques and chemical modification. Expression of the bactericidal peptide in Escherichia coli was accomplished through the formation of a fusion protein. The 5' end of the L-ribulokinase gene was fused to a single copy of a synthetic gene encoding cecropin A. A methionine codon was engineered between the two genes, and a methionylglycine extension was introduced at the C terminus of cecropin A. Cyanogen bromide treatment of the fusion protein yielded cecropin A with a C-terminal homoserine. The recombinant cecropin A with a homoserine at the C terminus did not kill most gram-positive bacteria tested. However, recombinant cecropin A with a chemically modified C terminus has antimicrobial activity similar to that of cecropin produced by cecropia pupae.

Molecular insight into mechanism of antimicrobial action of the β‐hairpin peptide arenicin: Specific oligomerization in detergent micelles

“…It cannot be said in advance which carrier protein will be suited well for this task in the case of each specific AMP. Incomplete list of the carrier proteins tested for this purpose includes: L-ribulokinase fragment, 15 glutathione-S-transferase, 16 Pseudomonas aeruginosa outer membrane protein, 16 Staphylococcus aureus protein A, and the dimer of its IgG-binding domain, 17 subtilisin inhibitor, 18 b-galactosidase, 18 maltose-binding protein, 18 prochymosin, 19 E.coli replication protein RepA and its fragment, 20 cellulose-binding domains, 20 human defensin HNP-1 preprodomain, 20 modified magainin intervening sequence (MIS), 21 GABA-transaminase, 22 truncated E.coli amidophosphoribosyltransferase, 23 Pseudomonas testosteroni ketosteroid isomerase, 24 fluorescent proteins GFP and obelin, 25 31 and ubiquitin. 32 An intein-based expression and purification system was also described.…”

Activity of the De Novo Engineered Antimicrobial Peptide WLBU2 against Pseudomonas aeruginosa in Human Serum and Whole Blood: Implications for Systemic Applications

“…These ubiquitous peptides typically consist of 20 to 40 amino acid residues and are highly cationic. Most host-derived CAPs display broad activity against both gram-positive and gramnegative bacteria (3,4,6,8,13).…”

Facilitation of Expression and Purification of an Antimicrobial Peptide by Fusion with Baculoviral Polyhedrin in Escherichia coli

“…Alternative expression strategies have been developed by the fusion of AMPs with partner proteins that decrease product toxicity to host cells, enhance product stability, facilitate product recovery, and/or aid in the acquisition of biological activity of the AMP product (5,6,10,11,16,25,26,29,36,39). Previous studies have investigated AMPs fused with several partners, including bovine prochymosin (11), maltose-binding protein (10,25), F4 of the E. coli intracellular protein PurF (16,28), green fluorescent protein (GFP) (36), Pap3.30 from the Pseudomonas aeruginosa bacteriophage PaP3 (29), bacterial thioredoxin (2), RepA of E. coli (39), intein (21), L-ribulokinase of Salmonella enterica serovar Typhimurium (5), the Cterminal fragment of light meromyosin (6), glutathione S-transferase (26), and immunoglobulin G (IgG)-binding domains from protein A (26).…”