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Molecular and Cellular Biochemistry

, Volume 140, Issue 1, pp 37–48 | Cite as

Membrane associated proteoglycans in rat testicular peritubular cells

  • Lahcen Bichoualne
  • Bénédicte Thiébot
  • Monique Langris
  • Pierre Barbey
  • Hamid Oulhaj
  • Jean Bocquet
Article

Abstract

Confluent testicular peritubular cells derived from immature rats were used to study membrane associated proteoglycans (PG) Peripheral material (heparin releasable), membrane and intracellular material (Triton X-100 releasable) were collected, purified by anion exchange chromatography then characterized by gel filtration and by hydrophobic interaction chromatography, followed by enzymatic digestion and chemical treatment. The peripheral material was constituted of two populations of PG (Kav=0 and 0.10 on Superose 6 column), each containing both heparan sulfate proteoglycans (HSPG) and chondroitin proteoglycans (CSPG) and perhaps a hybrid PG (HSCSPG). These PG being not retained on an octyl Sepharose column they were devoided of hydrophobic properties. The integral membrane proteoglycans isolated on the basis of their hydrophobic properties represented 20% of the Triton X-100 releasable material, and were exclusively constituted of proteoheparan sulfate. There were no relationships between this membrane HSPG and the peripheral HSPG as evidenced by pulse chase experiments. The mode of intercalation of the hydrophobic HSPG in the cell membrane was studied. The majority of these macromolecules (80%) were sensitive to trypsin and only a minor proportion (20%) were sensitive to phosphatidylinositol specific phospholipase C. Thus, about 80% of the hydrophobic HSPG were intercalated in the cell membrane by a hydrophobic segment of the core protein whereas about 20% were associated with the cell membrane via a phosphatidylinositol residue covalently bound to the core protein of the PG.

Key words

heparan sulfate proteoglycans rat testicular peritubular cells membrane associated proteoglycans 

Abbreviations

PG

Proteoglycans

CSPG

Chondroitin Sulfate Proteoglycans

HSPG

Heparan Sulfate Proteoglycans

HSCSPG

Heparan and Chondroitin Sulfate Proteoglycans

DNAse I

Deoxyribonuclease I

DMEM

Dulbeccos modified Eagle's medium

H/D

HAM F12/DMEM

ECM

Extracellular Matrix

PBS

Phosphate Buffered Saline

PI

Phosphatidylinositol

GPI

Glycosyl Phosphatidylinositol

PI-PLC

Phosphatidylinositol Specific Phospholipase C

TBS

Tris Buffered Saline

STI

Soybean Trypsin Inhibitor

GAG

Glycosaminoglycans

HA

Hyaluronic Acid

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Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • Lahcen Bichoualne
    • 1
  • Bénédicte Thiébot
    • 1
  • Monique Langris
    • 1
  • Pierre Barbey
    • 1
  • Hamid Oulhaj
    • 1
  • Jean Bocquet
    • 1
  1. 1.Laboratoire de Biochimie, ER CNRS 90, IRBAUniversité de CaenCAEN CedexFrance

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