title: Functional characterization of a short peptidoglycan recognition protein, PGRP5 in grass carp Ctenopharyngodon idella.
abstract: Peptidoglycan recognition proteins (PGRPs), which are evolutionarily conserved from insects to mammals, recognize bacterial peptidoglycan (PGN) and function in antibacterial innate immunity. In this study, a short-form PGRP, designated as gcPGRP5 was identified from grass carp Ctenopharyngodon idella. The deduced amino acid sequence of gcPGRP5 is composed of 180 residues with a conserved PGRP domain at the C-terminus. The gcPGRP5 gene consists of four exons and three introns, spacing approximately 2.3 kb in genomic sequence. Phylogenetic analysis demonstrated that the gcPGRP5 is clustered with other PGRP-S identified in teleost fish. The gcPGRP5 is constitutively expressed in all organs/tissues examined, and its expression was significantly induced in CIK cells treated with lipoteichoic acid (LTA), polyinosinic polycytidylic acid (Poly I:C) and PGN. Fluorescence analysis showed that gcPGRP5 is distributed in cytoplasm of CIK cells, and cell lysates from CIK cells transfected with pTurbo-gcPGRP5-GFP and ptGFP1-gcPGRP5 plasmids display the binding activity and peptidoglycan-lytic amidase activity toward Lys-PGN from Staphylococcus aureus and Dap-PGN from Bacillus subtilis. Furthermore, heat-shock protein70 (Hsp70), and MyD88, an adaptor molecule in Toll-like receptor pathway, had an increased expression in CIK cells overexpressed with gcPGRP5. It is thus indicated that gcPGRP5 exhibits amidase activity, and also possesses roles in anti-stress, and in Toll-like receptor signaling pathway.
authors: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei Province 430072, China.
authors: Jun Hua
authors: JH
authors: Li
authors: Ming Xian
authors: MX
authors: Chang
authors: Na Na
authors: NN
authors: Xue
authors: P
authors: P
authors: Nie
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001412:Bacillus subtilis / Q000502:physiology; D001483:Base Sequence; D002347:Carps / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism / Q000494:pharmacology; D016982:Micrococcus luteus / Q000502:physiology; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010457:Peptidoglycan / Q000378:metabolism / Q000494:pharmacology; D010802:Phylogeny; D011070:Poly I-C / Q000378:metabolism / Q000494:pharmacology; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D016415:Sequence Alignment / Q000662:veterinary; D013211:Staphylococcus aureus / Q000502:physiology; D013682:Teichoic Acids / Q000378:metabolism / Q000494:pharmacology
chemical_list: D002352:Carrier Proteins; D018076:DNA, Complementary; D029941:Fish Proteins; D006360:Heat-Shock Proteins; D008070:Lipopolysaccharides; D010457:Peptidoglycan; D012333:RNA, Messenger; D011994:Recombinant Proteins; D013682:Teichoic Acids; C118897:peptidoglycan recognition protein; C009900:lipoteichoic acid; D000581:Amidohydrolases; C019665:amidase; D011070:Poly I-C
keywords: Amidase activity; Grass carp; PGRP5; Peptidoglycan recognition protein; Peptidoglycan-binding activity