<var date-time="ALnxMWdz"></var> VSports最新版本 - Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The . gov means it’s official. Federal government websites often end in . gov or . mil VSports app下载. Before sharing sensitive information, make sure you’re on a federal government site. .

Https

The site is secure. The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely V体育官网. .

. 2005 Oct 1;106(7):2551-8.
doi: 10.1182/blood-2005-02-0530. Epub 2005 Jun 14.

Human peptidoglycan recognition protein S is an effector of neutrophil-mediated innate immunity

Ju Hyun Cho  1 , Iain P Fraser, Koichi Fukase, Shoichi Kusumoto, Yukari Fujimoto, Gregory L Stahl, R Alan B Ezekowitz
Affiliations

"V体育2025版" Human peptidoglycan recognition protein S is an effector of neutrophil-mediated innate immunity

Ju Hyun Cho et al. Blood. .

Abstract

Innate immune responses to bacteria require cooperative interactions between host recognition molecules and phagocytes. The peptidoglycan recognition proteins (PGRPs) are a large group of proteins found in insects and mammals that bind to bacterial peptidoglycan (PGN). PGRP-S is located with other antimicrobial proteins, such as lysozyme, in the granules of human neutrophils. Whereas both PGRP-S and lysozyme recognize PGN, the exact binding specificity of human PGRP-S, its functional activity, and its potential synergy with other neutrophil-derived bactericidal proteins such as lysozyme have not been determined. Here we show that human PGRP-S binds to and inhibits the growth of Staphylococcus aureus (containing lysine-type PGN) and Escherichia coli (containing mesodiaminopimelic acid-type PGN). The binding affinity and thus antimicrobial activity of PGRP-S is determined by the third amino acid in the PGN stem peptide. Furthermore, the antimicrobial effect of PGRP-S against E coli is synergistic with lysozyme, and lysozyme and PGRP-S colocalize in neutrophil extracellular traps (NETs), suggesting that these granule-derived proteins act together to kill bacteria trapped in the NETs VSports手机版. Taken together, these results indicate that human PGRP-S plays a role in innate immunity in the context of neutrophils by contributing to the killing of intracellular and extracellular bacteria. .

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Expression and purification of recombinant human PGRP-S. (A) Cell culture supernatant and cell lysates were tested for expression by Western blot with anti-His(C-term) antibody. (B) Coomassie Blue-stained 12% SDS-PAGE gel showing affinity-purified rhPGRP-S. Molecular markers are shown on the left. (C) Gel filtration chromatogram showing elution of rhPGRP-S from a Superdex 75HR column at a flow rate of 0.5 mL/min. Based on elution volumes of molecular weight standards, rhPGRP-S elutes as a monomer of approximately 23 kDa. (D) fast protein liquid chromatography (FPLC) fractions containing rhPGRP-S were subjected to 12% SDS-PAGE.
Figure 2.
Figure 2.
Human PGRP-S binds to S aureus CP5 and E coli K12. Bacteria (2 × 106) were incubated with biotin-labeled rhPGRP-S (10 μg/mL) and BSA (40 μg/mL), respectively, at 37°C for 30 minutes and visualized with streptavidin-Alexa Fluor 488.
Figure 3.
Figure 3.
Human PGRP-S binds to Lys-type and DAP-type PGNs. Purified rhPGRP-S or rdPGRP-LC (0.5 μg) were incubated with insoluble Lys-type or DAP-type PGN, and bound protein on the insoluble PGN was separated from unbound protein, as described in “Materials and methods.” One tenth of unbound protein (lane U) and one fifth of bound protein (lane B) were analyzed by Western blot analysis using anti-His(C-term) antibody. Molecular markers are indicated on the left.
Figure 4.
Figure 4.
Human PGRP-S inhibits the growth of S aureus CP5 and E coli K12. (A) Radial diffusion assay. Wells were bored into underlay gel (1% agarose in 0.01 × TSB, 10 mM NaPB, pH 7.4) impregnated with bacteria. Proteins were introduced into wells (0.1-5 μg/well), and the plates were overlaid with 1% agarose in 2 × TSB (6% wt/vol) and incubated overnight at 37°C. (B) Suspension assay. Bacteria were incubated in 1 × TSB (3% wt/vol) with 25 μg/mL rhPGRP-S, either alone (•) or supplemented with 100 μg/mL S aureus PGN or E coli PGN (▴), PGN only (formula image) or no additives (⋄). Tubes were shaken at 300 rpm for 5 hours, and bacterial density was monitored by measurement of optic density (OD) at 600 nm at 1-hour intervals. Data represent the mean ± SD of 3 independent experiments.
Figure 5.
Figure 5.
Human PGRP-S shows synergistic antibacterial effect with lysozyme against E coli K12. (A) Lysis of lysozyme-treated E coli cells. Lysozyme-treated E coli cells (⋄) were incubated in 10 mM NaPB, pH 7.4, with rhPGRP-S (4 μg/mL, [▴]; 8 μg/mL, [•]) or PMBN (2.5 μg/mL, [formula image]), and the change in OD at 600 nm was monitored for 10 minutes. Data represent the mean ± SD of 3 independent experiments. (B) Growth inhibition of E coli cells. E coli cells were incubated in 1 × TSB (3% wt/vol) with 50 μg/mL lysozyme (formula image), 25 μg/mL rhPGRP-S (▴), 8 μg/mL rhPGRP-S and 50 μg/mL of lysozyme (•), or no protein (⋄). Tubes were shaken at 300 rpm for 5 hours, and bacterial density was monitored by measurement of optic density (OD) at 600 nm at 1-hour intervals. Data represent the mean of duplicate samples from 1 of 2 similar experiments.
Figure 6.
Figure 6.
Colocalization of PGRP-S with lysozyme in the NETs. Neutrophils were activated with 100 nM PMA for 30 minutes and stained for DNA (A), PGRP-S (B), and lysozyme (C). For DNA detection, Hoechst 33342 (blue) was used. PGRP-S was detected with anti-human PGRP-S monoclonal antibody (mouse immunoglobulin G [IgG]) and Cy2-conjugated donkey anti-mouse IgG (green). Lysozyme was detected with anti-human lysozyme polyclonal antibody (sheep IgG) and Cy5-conjugated donkey anti-sheep IgG (red). The merged image (D) clearly shows colocalization of PGRP-S with lysozyme in the NETs.
See this image and copyright information in PMC

References

    1. Hoffmann JA, Kafatos FC, Janeway CA, Ezekowitz RAB. Phylogenetic perspectives in innate immunity. Science. 1999;284: 1313-1318. - PubMed
    1. Janeway CA, Medzhitov R. Innate immune recognition. Annu Rev Immunol. 2002;20: 197-216. - PubMed
    1. Hoffmann JA. The immune response of Drosophila. Nature. 2003;426: 33-38. - PubMed (V体育ios版)
    1. Yoshida H, Kinoshita K, Ashida M. Purification of a peptidoglycan recognition protein from hemolymph of the silkworm, Bombyx mori. J Biol Chem. 1996;271: 13854-13860. - PubMed
    1. Ochiai M, Ashida M. A pattern recognition protein for peptidoglycan: cloning the cDNA and the gene of the silkworm, Bombyx mori. J Biol Chem. 1999;274: 11854-11858. - VSports在线直播 - PubMed

Publication types

MeSH terms (V体育官网)