A prawn core histone 4: Derivation of N- and C-terminal peptides and their antimicrobial properties, molecular characterization and mRNA transcription

Abstract

This study investigates the complete molecular characterization including bioinformatics characterization, gene expression, synthesis of N and C terminal peptides and their antimicrobial activity of the core histone 4 (H4) from freshwater giant prawn Macrobrachium rosenbergii (Mr). A cDNA encoding MrH4 was identified from the constructed cDNA library of M. rosenbergii during screening and the sequence was obtained using internal sequencing primers. The MrH4 coding region possesses a polypeptide of 103 amino acids with a calculated molecular weight of 11kDa and an isoelectric point of 11.5. The bioinformatics analysis showed that the MrH4 polypeptide contains a H4 signature at (15)GAKRH(19). Multiple sequence alignment of MrH4 showed that the N-terminal (21-42) and C-terminal (87-101) antimicrobial peptide regions and the pentapeptide or H4 signature (15-19) are highly conserved including in humans. The phylogenetic tree formed two separate clades of vertebrate and invertebrate H4, wherein MrH4 was located within the arthropod monophyletic clade of invertebrate H4 groups. Three-dimensional model of MrH4 was established using I-TASSER program and the model was validated using Ramachandran plot analysis. Schiffer-Edmundson helical wheel modeling was used to predict the helix propensity of N (21-42) and C (87-101) terminal derived Mr peptides. The highest gene expression was observed in gills and is induced by viral [white spot syndrome baculovirus (WSBV) and M. rosenbergii nodovirus (MrNV)] and bacterial (Aeromonas hydrophila and Vibrio harveyi) infections. The N and C terminal peptides were synthesized and their antimicrobial and hemolytic properties were examined. Both peptides showed activity against the tested Gram negative and Gram positive bacteria; however, the highest activity was noticed against Gram negative bacteria. Among the two peptides used in this study, C-terminal peptide yielded better results than the N-terminal peptide. Therefore, C terminal peptide can be recommended for the development of an antimicrobial agent.

Impacts of environmental and biological stressors on immune system ofMacrobrachium rosenbergii

“…; Chaurasia et al . b), anti‐lipopolysaccharide factor (Arockiaraj et al . ) and lectin (Arockiaraj et al .…”

“…Similarly in another study, Chaurasia et al . (,b) have reported an 11 kDa histone 4 protein with a penta‐peptide H4 signature where two antimicrobial peptide regions were derived from histone 4. Both the peptides (N‐terminal: 21 KVLRDNIQGITKPAIRRLARRG 42 and C‐terminal: 87 VVYALKRQGRTLYGF 101 ) showed better inhibition against Gram‐negative than the Gram‐positive bacteria.…”

Proteomic and structural differences in lumpfish skin among the dorsal, caudal and ventral regions

“…rosenbergii noda virus, A . hydrophila and Vibrio harveyi 34 . Histone proteins have been identified in skin/skin mucus of several teleosts such as histone H4 in mrigal ( Cirrhinus mrigala ) 35 and European sea bass ( D .…”

Antimicrobial peptides of invertebrates. Part 1. structure, biosynthesis, and evolution

“…Among AMPs of invertebrates, derivatives of hemocy anin [174,175] and histones [176,177] belong to this group. These proteins can be considered as the AMP depots in tissues which allow fast immobilization of protective resources of an organism through activation of specific proteases [174] or via nonenzymatic frag mentation [178].…”