Crustin, a WAP domain containing antimicrobial peptide from freshwater prawn Macrobrachium rosenbergii: immune characterization.
Crustin (MrCrs) was sequenced from a freshwater prawn Macrobrachium rosenbergii. The MrCrs protein contains a signal peptide region at N-terminus between 1 and 22 and a long whey acidic protein domain (WAP domain) at C-terminus between 57 and 110 along with a WAP-type 'four-disulfide core' motif. Phylogenetic results show that MrCrs is clustered together with other crustacean crustin groups. MrCrs showed high sequence similarity (77%) with crustin from Pacific white shrimp Litopenaeus vannamei and Japanese spiny lobster Panulirus japonicas. I-TASSER uses the best structure templates to predict the possible structures of MrCrs along with PDB IDs such as 2RELA and 1FLEI. The gene expressions of MrCrs in both healthy M. rosenbergii and those infected with virus including infectious hypodermal and hematopoietic necrosis virus (IHHNV) and white spot syndrome virus (WSSV) and bacteria Aeromonas hydrophila (Gram-negative) and Enterococcus faecium (Gram-positive) were examined using quantitative real time PCR. To understand its biological activity, the recombinant MrCrs gene was constructed and expressed in Escherichia coli BL21 (DE3). The recombinant MrCrs protein agglutinated with the bacteria considered for analysis at a concentration of 25 μg/ml, except Lactococcus lactis. The bactericidal results showed that the recombinant MrCrs protein destroyed all the bacteria after incubation, even less than 6 h. These results suggest that MrCrs is a potential antimicrobial peptide, which is involved in the defense system of M. rosenbergii against viral and bacterial infections.
Department of Biotechnology, Faculty of Science and Humanities, SRM University, SRM Nagar, Kattankulathur, Chennai, Tamil Nadu, India. jesuaraj@gmail.com
Jesu
J
Arockiaraj
Annie J
AJ
Gnanam
Dhanaraj
D
Muthukrishnan
Ranganath
R
Gudimella
James
J
Milton
Arun
A
Singh
Saravanan
S
Muthupandian
Marimuthu
M
Kasi
Subha
S
Bhassu
D016980:Aeromonas hydrophila / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D060829:Arthropod Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D019223:Densovirinae / Q000502:physiology; D016984:Enterococcus faecium / Q000502:physiology; D020869:Gene Expression Profiling / Q000662:veterinary; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D009928:Organ Specificity; D033541:Palaemonidae / Q000235:genetics* / Q000276:immunology* / Q000382:microbiology / Q000821:virology; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment / Q000662:veterinary; D046848:White spot syndrome virus 1 / Q000502:physiology
D023181:Antimicrobial Cationic Peptides; D060829:Arthropod Proteins; D018076:DNA, Complementary; D012333:RNA, Messenger