Association of laminin with heparan and chondroitin sulfate-bearing proteoglycans in neurite-promoting factor complexes from rat schwannoma cells

Neurochem Res. 1987 Oct;12(10):909-21. doi: 10.1007/BF00966313.

Abstract

The present studies were undertaken to confirm the presence and identity of a putative proteoglycan associated with laminin in neurite-promoting factor complexes isolated from rat schwannoma cell conditioned medium. Sucrose density gradient centrifugation of the complex resolved two laminin-associated Na2[35S]O4-labeled peaks which were termed Pools A and B. Both pools had nearly all their [35S] cpms associated with glycosaminoglycan, contained heparan sulfate-proteoglycan core protein antigen and displayed a similarly high neurite promoting potency relative to their laminin contents. However, Pool A contained about twice as many [35S] cpms and twice as much proteoglycan core protein per laminin than Pool B. Seventy percent of Pool A cpms was associated with heparan sulfate and 30% with chondroitin sulfate whereas the inverse was true for Pool B. Treatment with heparitinase and/or chondroitinase ABC caused laminin in either pool to elute at lower salt concentrations from DEAE cellulose. In SDS-PAGE the [35S] cpms of both pools ran with the same mobility as laminin but could be separated from laminin under reducing conditions. The Pool A cpms remained at 900 KD and the Pool B cpms spread over the 200-900 KD range. By rotary shadowing electron microscopy, Pool B fractions contained primarily cross-shaped laminin images, often associated with proteoglycan-like images. Pool A fractions contained i) dense, aggregated images including intact laminin from which emanated proteoglycan-like strands, ii) circular images bearing globular domains and less commonly, iii) distorted cross-shaped laminin-like images. These studies support the existence of at least two forms of laminin-proteoglycan complexes which differ in biochemical, immunochemical and ultrastructural characteristics.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cells, Cultured
  • Chondroitin / analogs & derivatives*
  • Chondroitin Sulfates / analysis*
  • Glycosaminoglycans / analysis*
  • Heparitin Sulfate / analysis*
  • Laminin / analysis*
  • Nerve Growth Factors*
  • Neurilemmoma / analysis*
  • Peptides / analysis
  • Peptides / isolation & purification*
  • Proteoglycans / analysis*
  • Rats

Substances

  • Glycosaminoglycans
  • Laminin
  • Nerve Growth Factors
  • Peptides
  • Proteoglycans
  • neurite-inducing factor
  • Chondroitin
  • Chondroitin Sulfates
  • Heparitin Sulfate