title: The non-aggregated aggrecan in the human intervertebral disc can arise by a non-proteolytic mechanism.
abstract: Analysis of both the aggregated and non-aggregated fractions of aggrecan isolated from adult human intervertebral disc using immunoblotting with antibodies specific for the different domains constituting the aggrecan core protein or atomic force microscopy revealed that many components contained the G1 domain. However, little of the disc aggrecan was able to reform aggregates with hyaluronan, as determined by gel filtration chromatography, suggesting that the G1 domains had been rendered non-functional. Since previous studies have shown that disc aggrecan undergoes non-enzymatic glycation with age, the functional effect of such modification was investigated in vitro using bovine aggrecan isolated from young animals. Incubation of monomeric aggrecan with ribose to induce glycation rendered it unable to form complexes with hyaluronan stable to agarose gel electrophoresis or gel filtration chromatography. Similarly, extended treatment of intact proteoglycan aggregate with ribose resulted in destabilisation of the complex with separation of the aggrecan from the hyaluronan. Although it is clear that proteolysis occurs in the intervertebral disc and gives rise to some non-aggregating molecules, a different mechanism is required to explain the presence of many non-aggregating molecules bearing the G1 domain. The products of non-enzymatic glycation of the globular domains of aggrecan would account for this phenomenon and explain why some of the non-aggregating molecules are still large proteoglycans. While such molecules may be retained in the nucleus pulposus, they may be able to diffuse within it, reducing the ability of the tissue to resist compression under asymmetric loading such as bending and ultimately contributing to disc degeneration.
authors.affiliation: Genetics Unit, Shriners Hospital for Children, 1529 Cedar Avenue, Montreal, Quebec H3G 1A6, Canada.jmort@shriners.mcgill.ca.
authors.forename: P J
authors.initials: PJ
authors.lastname: Roughley
authors.forename: Y
authors.initials: Y
authors.lastname: Geng
authors.forename: J S
authors.initials: JS
authors.lastname: Mort
mesh_terms: D000328:Adult; D053674:Aggrecans / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D006801:Humans; D007403:Intervertebral Disc / Q000378:metabolism*; D066329:Protein Aggregates; D059748:Proteolysis
chemical_list: D053674:Aggrecans; D066329:Protein Aggregates