Demonstration of peptidoglycan-binding sites on lymphocytes and macrophages by photoaffinity cross-linking

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

One dominant binding site (70 kDa 6.5 pI protein) for bacterial cell wall peptidoglycan (PGN), a macrophage activator and polyclonal B cell mitogen, was demonstrated on mouse B and T lymphocytes and macrophages by photoaffinity cross-linking and two-dimensional polyacrylamide gel electrophoresis. This binding site was not present on erythrocytes. The binding was specific for polymeric PGN and was competitively inhibited by unlabeled PGN with IC50 = 48 μg/ ml (0.38 μm). The binding was partially inhibited by O-acetylated PGN monomers (IC50 = 469 μg/ml, 521 μM), dextran sulfate (IC50 = 1024 μg/ml, 124 μM), and (GlcNAc)3 (IC50 = 6.6 mg/ml, 10 mM), and was not inhibited by non-O-acetylated PGN monomers and dimers, muramyl dipeptide, PGN pentapeptide, GlcNAc, teichoic acid, protein A, and gelatin. The cell surface location of the 70-kDa PGN-binding protein was indicated by the ability of PGN to bind to this protein in intact metabolically inactive cells (at 4°C and in the presence of 0.1% NaN3) and by the ability to extract the 70-kDa PGN-binding protein from viable B lymphocytes by noncytotoxic concentration of n-octyl-β-D-glucopyranoside.

Original languageEnglish
Pages (from-to)4713-4718
Number of pages6
JournalJournal of Biological Chemistry
Volume266
Issue number8
StatePublished - Mar 15 1991

Fingerprint

Lymphocytes
Peptidoglycan
Macrophages
Demonstrations
Binding Sites
Inhibitory Concentration 50
Carrier Proteins
B-Lymphocytes
Monomers
Teichoic Acids
Acetylmuramyl-Alanyl-Isoglutamine
Sodium Azide
Dextran Sulfate
Bacterial Proteins
T-cells
Electrophoresis, Gel, Two-Dimensional
Staphylococcal Protein A
Gelatin
Electrophoresis
Mitogens

ASJC Scopus subject areas

  • Biochemistry

Cite this

Demonstration of peptidoglycan-binding sites on lymphocytes and macrophages by photoaffinity cross-linking. / Dziarski, Roman.

In: Journal of Biological Chemistry, Vol. 266, No. 8, 15.03.1991, p. 4713-4718.

Research output: Contribution to journalArticle

@article{280aebee29da4206aa6dde7209b00f1d,
title = "Demonstration of peptidoglycan-binding sites on lymphocytes and macrophages by photoaffinity cross-linking",
abstract = "One dominant binding site (70 kDa 6.5 pI protein) for bacterial cell wall peptidoglycan (PGN), a macrophage activator and polyclonal B cell mitogen, was demonstrated on mouse B and T lymphocytes and macrophages by photoaffinity cross-linking and two-dimensional polyacrylamide gel electrophoresis. This binding site was not present on erythrocytes. The binding was specific for polymeric PGN and was competitively inhibited by unlabeled PGN with IC50 = 48 μg/ ml (0.38 μm). The binding was partially inhibited by O-acetylated PGN monomers (IC50 = 469 μg/ml, 521 μM), dextran sulfate (IC50 = 1024 μg/ml, 124 μM), and (GlcNAc)3 (IC50 = 6.6 mg/ml, 10 mM), and was not inhibited by non-O-acetylated PGN monomers and dimers, muramyl dipeptide, PGN pentapeptide, GlcNAc, teichoic acid, protein A, and gelatin. The cell surface location of the 70-kDa PGN-binding protein was indicated by the ability of PGN to bind to this protein in intact metabolically inactive cells (at 4°C and in the presence of 0.1{\%} NaN3) and by the ability to extract the 70-kDa PGN-binding protein from viable B lymphocytes by noncytotoxic concentration of n-octyl-β-D-glucopyranoside.",
author = "Roman Dziarski",
year = "1991",
month = "3",
day = "15",
language = "English",
volume = "266",
pages = "4713--4718",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "8",

}

TY - JOUR

T1 - Demonstration of peptidoglycan-binding sites on lymphocytes and macrophages by photoaffinity cross-linking

AU - Dziarski, Roman

PY - 1991/3/15

Y1 - 1991/3/15

N2 - One dominant binding site (70 kDa 6.5 pI protein) for bacterial cell wall peptidoglycan (PGN), a macrophage activator and polyclonal B cell mitogen, was demonstrated on mouse B and T lymphocytes and macrophages by photoaffinity cross-linking and two-dimensional polyacrylamide gel electrophoresis. This binding site was not present on erythrocytes. The binding was specific for polymeric PGN and was competitively inhibited by unlabeled PGN with IC50 = 48 μg/ ml (0.38 μm). The binding was partially inhibited by O-acetylated PGN monomers (IC50 = 469 μg/ml, 521 μM), dextran sulfate (IC50 = 1024 μg/ml, 124 μM), and (GlcNAc)3 (IC50 = 6.6 mg/ml, 10 mM), and was not inhibited by non-O-acetylated PGN monomers and dimers, muramyl dipeptide, PGN pentapeptide, GlcNAc, teichoic acid, protein A, and gelatin. The cell surface location of the 70-kDa PGN-binding protein was indicated by the ability of PGN to bind to this protein in intact metabolically inactive cells (at 4°C and in the presence of 0.1% NaN3) and by the ability to extract the 70-kDa PGN-binding protein from viable B lymphocytes by noncytotoxic concentration of n-octyl-β-D-glucopyranoside.

AB - One dominant binding site (70 kDa 6.5 pI protein) for bacterial cell wall peptidoglycan (PGN), a macrophage activator and polyclonal B cell mitogen, was demonstrated on mouse B and T lymphocytes and macrophages by photoaffinity cross-linking and two-dimensional polyacrylamide gel electrophoresis. This binding site was not present on erythrocytes. The binding was specific for polymeric PGN and was competitively inhibited by unlabeled PGN with IC50 = 48 μg/ ml (0.38 μm). The binding was partially inhibited by O-acetylated PGN monomers (IC50 = 469 μg/ml, 521 μM), dextran sulfate (IC50 = 1024 μg/ml, 124 μM), and (GlcNAc)3 (IC50 = 6.6 mg/ml, 10 mM), and was not inhibited by non-O-acetylated PGN monomers and dimers, muramyl dipeptide, PGN pentapeptide, GlcNAc, teichoic acid, protein A, and gelatin. The cell surface location of the 70-kDa PGN-binding protein was indicated by the ability of PGN to bind to this protein in intact metabolically inactive cells (at 4°C and in the presence of 0.1% NaN3) and by the ability to extract the 70-kDa PGN-binding protein from viable B lymphocytes by noncytotoxic concentration of n-octyl-β-D-glucopyranoside.

UR - http://www.scopus.com/inward/record.url?scp=0025855140&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025855140&partnerID=8YFLogxK

M3 - Article

C2 - 2002020

AN - SCOPUS:0025855140

VL - 266

SP - 4713

EP - 4718

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 8

ER -