title: The N-terminal glycine-rich and cysteine-rich regions are essential for antimicrobial activity of crustinPm1 from the black tiger shrimp Penaeus monodon.
abstract: An antimicrobial protein crustinPm1 from Penaeus monodon is a WAP domain-containing protein with an antimicrobial activity against Gram-positive bacteria but does not have antiproteinase activity. The lack of antiproteinase is speculated to be due to the P(1)' Met and/or the length of spacing between the conserved Cys2 and Cys3 while the antimicrobial activity may be due to the N-terminal Gly-rich and Cys-rich regions. In this study, the P(1)-P(1)' and the N-terminal Gly-rich and Cys-rich regions of crustinPm1 were mutated by amino acid substitution or deletion. Substitutions of P(1)-P(1)' from Pro-Pro to Leu-Leu, Leu-His, Leu-Met, Leu-Ala and P(1)' from Pro to Met did not make the protein inhibitory to subtilisin, trypsin, chymotrypsin and elastase. The mutations at P(1)-P(1)' positions in rcrustinPm1 had no effect on antibacterial activity. The WAP domain mutant with both Gly-rich and Cys-rich regions deleted did not exhibit antibacterial activity against Staphylococcus aureus while the deletion mutants of either Gly-rich or Cys-rich regions exhibited lower antibacterial activity than the wild type crustinPm1. Therefore, both Gly-rich and Cys-rich regions attached to a WAP domain are essential for efficient antibacterial activity of crustinPm1.
authors.affiliation: Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Phyathai Road, Pathumwan, Bangkok 10330, Thailand.
authors.forename: Pranisa
authors.initials: P
authors.lastname: Suthianthong
authors.forename: Suchao
authors.initials: S
authors.lastname: Donpudsa
authors.forename: Premruethai
authors.initials: P
authors.lastname: Supungul
authors.forename: Anchalee
authors.initials: A
authors.lastname: Tassanakajon
authors.forename: Vichien
authors.initials: V
authors.lastname: Rimphanitchayakit
mesh_terms: D020816:Amino Acid Motifs; D019943:Amino Acid Substitution; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D003545:Cysteine / Q000235:genetics; D020869:Gene Expression Profiling; D005998:Glycine / Q000235:genetics; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D033561:Penaeidae / Q000235:genetics* / Q000378:metabolism* / Q000382:microbiology; D010447:Peptide Hydrolases / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment; D013211:Staphylococcus aureus / Q000502:physiology
chemical_list: D023181:Antimicrobial Cationic Peptides; D050505:Mutant Proteins; D011994:Recombinant Proteins; D010447:Peptide Hydrolases; D003545:Cysteine; D005998:Glycine