The serine proteinase inhibitory proteins of the chondrodystrophoid (beagle) and non-chondrodystrophoid (greyhound) canine intervertebral disc.
Trypsin inhibitory proteins of low buoyant density (p < or = 1.35 g/mL) fractions were prepared by CsCl density gradient ultracentrifugation of 4 M guanidinium hydrochloride extracts of lumbar beagle and greyhound annulus fibrosus and nucleus pulposus from animals aged 1 to 6 years. Affinity blotting with biotinylated trypsin was used to identify active trypsin inhibitory protein species; these species were also identified immunologically by Western blotting using antibodies against bovine pancreatic trypsin inhibitor (BPTI), and human inter-alpha-trypsin inhibitor (ITI). None of the trypsin inhibitory species evident in Western blots were reactive with anti-human alpha1-proteinase inhibitor (alpha-1-PI), alpha2-macroglobulin or secretory leucocyte proteinase inhibitor. The greyhound intervertebral disc samples generally had higher levels of active trypsin inhibitor species per unit weight of tissue extracted, and a more extensive range of inhibitor species. Inhibitor species of 30, 32, 34 kDa were identified in both beagle and greyhound intervertebral disc samples; these species were generally most prominent in the annulus fibrosus samples. In contrast, the nucleus pulposus samples contained relatively large trypsin inhibitor species; the anti-BPTI detected an inhibitor species of approximately 85-90 kDa; anti-ITI detected species of 120-250 kDa; biotinylated trypsin detected species of 60-110 kDa. A small molecular mass trypsin inhibitor species of 6 kDa, which was of similar mobility to BPTI, was also detected in annulus fibrosus samples; however, this species did not react with anti-BPTI.
Melrose
J
J
Raymond Purves Bone and Joint Research Laboratories, The University of Sydney and The Royal North Shore Hospital of Sydney, St. Leonards, NSW, Australia.
Taylor
T K
TK
Ghosh
P
P
D017668:Age of Onset; D000818:Animals; D007611:Aprotinin / Q000032:analysis; D002417:Cattle; D004285:Dogs; D006801:Humans; D007403:Intervertebral Disc / Q000737:chemistry*; D011506:Proteins / Q000032:analysis*; D013045:Species Specificity; D013122:Spinal Diseases / Q000662:veterinary; D014361:Trypsin Inhibitors / Q000032:analysis*
D011506:Proteins; D014361:Trypsin Inhibitors; D007611:Aprotinin