PMID- 2447876
OWN - NLM
STAT- MEDLINE
DCOM- 19880204
LR  - 20200304
IS  - 0264-6021 (Print)
IS  - 1470-8728 (Electronic)
IS  - 0264-6021 (Linking)
VI  - 247
IP  - 3
DP  - 1987 Nov 1
TI  - Identification of a 64 kDa heparan sulphate proteoglycan core protein from human 
      lung fibroblast plasma membranes with a monoclonal antibody.
PG  - 765-71
AB  - Human lung fibroblasts produce heparan sulphate proteoglycans (HSPG) that are 
      associated with the plasma membrane. A monoclonal-antibody (Mab)-secreting 
      hybridoma, S1, was produced by fusion of SP 2/0-AG 14 mouse myeloma cells with 
      spleen cells from mice immunized with partially purified cellular HSPG fractions. 
      The HSPG character of the material carrying the epitope recognized by Mab S1 was 
      demonstrated by: (i) the co-purification of the S1 epitope with the membrane HSPG 
      of human lung fibroblasts; (ii) the decrease in size of the material carrying the 
      S1 epitope upon treatment with heparinase or heparitinase, and the resistance of 
      this material to heparinase treatment after N-desulphation. The S1 epitope 
      appears to be part of the core protein, since it was destroyed by proteinase 
      treatment and by disulphide-bond reduction, but not by treatments that 
      depolymerize the glycosaminoglycan chains and N-linked oligosaccharide chains. 
      Polyacrylamide-gel electrophoresis of non-reduced heparitinase-digested membrane 
      HSPG followed by Western blotting and immunostaining with Mab S1 revealed a 
      single band with apparent molecular mass of 64 kDa. Membrane proteoglycans 
      isolated from detergent extracts or from 4 M-guanidinium chloride extracts of the 
      cells yielded similar results. Additional digestion with N-glycanase lowered the 
      apparent molecular mass of the immunoreactive material to 56 kDa, suggesting that 
      the core protein also carries N-linked oligosaccharides. Fractionation of 
      125I-labelled membrane HSPG by immuno-affinity chromatography on immobilized Mab 
      S1, followed by heparitinase digestion and polyacrylamide-gel electrophoresis of 
      the bound material, yielded a single labelled band with apparent molecular mass 
      64 kDa. Treatment with dithiothreitol caused a slight increase in apparent 
      molecular mass, suggesting that the core protein of this membrane proteoglycan of 
      a single subunit containing (an) intrachain disulphide bond(s).
FAU - de Boeck, H
AU  - de Boeck H
AD  - Center for Human Genetics, University of Leuven, Belgium.
FAU - Lories, V
AU  - Lories V
FAU - David, G
AU  - David G
FAU - Cassiman, J J
AU  - Cassiman JJ
FAU - van den Berghe, H
AU  - van den Berghe H
LA  - eng
GR  - HL-31750/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Chondroitin Sulfate Proteoglycans)
RN  - 0 (Epitopes)
RN  - 0 (Glycosaminoglycans)
RN  - 0 (Heparan Sulfate Proteoglycans)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (Proteoglycans)
RN  - 9050-30-0 (Heparitin Sulfate)
SB  - IM
MH  - *Antibodies, Monoclonal
MH  - Cells, Cultured
MH  - Chondroitin Sulfate Proteoglycans/immunology/*isolation & purification
MH  - Chromatography, Gel
MH  - Epitopes/analysis
MH  - Fibroblasts/analysis
MH  - Glycosaminoglycans/*isolation & purification
MH  - Heparan Sulfate Proteoglycans
MH  - Heparitin Sulfate/immunology/*isolation & purification
MH  - Humans
MH  - Immunoelectrophoresis
MH  - Lung/*analysis
MH  - Membrane Glycoproteins/*isolation & purification
MH  - Proteoglycans/*isolation & purification
PMC - PMC1148477
EDAT- 1987/11/01 00:00
MHDA- 1987/11/01 00:01
PMCR- 1988/05/01
CRDT- 1987/11/01 00:00
PHST- 1987/11/01 00:00 [pubmed]
PHST- 1987/11/01 00:01 [medline]
PHST- 1987/11/01 00:00 [entrez]
PHST- 1988/05/01 00:00 [pmc-release]
AID - 10.1042/bj2470765 [doi]
PST - ppublish
SO  - Biochem J. 1987 Nov 1;247(3):765-71. doi: 10.1042/bj2470765.