Proteoglycan heterogeneity in the normal adult ovine intervertebral disc

Proteoglycans (PGs) were isolated from 4 M GuHCl extracts of young adult ovine Intervertebral disc (IVD) tissues using sequential CsCl density gradient centrifugation, and a combination of gel-permeation and hydrophobic chromatography. A total of six PG sub-populations were identified in both the Annulus fibrosus (AF) and Nucleus pulposus (NP), i.e. two high buoyant density aggregatable PGs, two high buoyant density non-aggregatable PGs and two small, low-intermediate buoyant density, non-aggregatable, DS-rich PG species. These latter PGs were identified as biglycan and decorin on the basis of analyses of their core protein native size, and glycosaminoglycan composition. Additional low-intermediate buoyant density PG species were also evident in the non-aggregatable PG pool, particularly in extracts of NP tissues. These PGs did not bind to the octyl affinity matrix under the experimental conditions employed and thus were readily separated from the DS-PGs by hydrophobic chromatography, their constituent glycosaminoglycans (CS and KS) also differed and were of a smaller size to the CS and KS chains isolated from the large high buoyant density PGs, the small PGs which did not bind to octyl-sepharose may therefore represent distinct PG species in their own right. Differences were evident in the absolute size, and in the distribution of individual PG species in the respective IVD tissues but the NP always contained a larger proportion of high buoyant density non-aggregatable PGs of somewhat smaller size than those isolated from the AF. Decorin and biglycan, however, were generally more abundant in AF tissues.