Recovery of bone and serum proteins 



from human skeletal tissue: IgG, 



osteonectin, and albumin 



Noreen Tuross 



The most common substrates from which to infer or deduce 

 vertebrate paleopathology are bones and teeth. Physical an- 

 thropologists have explored the significant contribution that 

 morphology and histology of mineralized tissues can make to 

 our understanding of paleopathology. This study describes 

 information at the molecular level that remains in the bones 

 and teeth of some of our ancestors. 



The presence of amino acids in fossil bones and teeth was 

 the first evidence that proteins indigenous to the animal 

 might remain in the mineralized tissues (Abelson 1956; Ho 

 1965). The preservation of the amino acids from the major 

 bone protein, collagen, was documented in a variety of fos- 

 sil bones from many locations (Ho 1965; Wyckoff and 

 Doberenz 1965; Tuross and Hare 1978). In fossil bones much 

 of the collagen exists as a heterogeneous mixture of degrada- 

 tion products relative to the original gene product (Tuross et 

 al. 1980). Isotopically. however, the degraded collagen is 

 assumed to retain the pertinent information for archeometric 

 use. Radiocarbon from collagen has been used to date fossils 

 (Libby 1955; Taylor 1987) and the stable isotopes of carbon 

 and nitrogen from bone collagen have been used in paleodic- 

 tary interpretations (Schoeninger et al. 1983; Schoeninger 

 and DeNiro 1984). Degradation of collagen, particularly the 

 relative loss of the amino acid, glycine, can perturb the car- 

 bon and nitrogen stable isotope values obtained from fossil 

 bones and teeth (Tuross et al . 1 988 ). Better methods of isola- 

 tion and characterization of proteins from fossil bones and 

 teeth will contribute to the accuracy of archeometric isotopic 

 applications. 



Many proteins other than collagen can be found in modern 

 bone. Both scrum-derived and bone-cell-produced proteins 

 can be extracted from bone. The complexity of the mixture of 

 proteins found in bone can be seen in a two-dimensional gel 

 electrophoresis analysis of bovine bone where approximately 

 2(X) separate proteins were observed (Dclmas et al. 1984). 



Extraction techniques developed by John Termine and 

 coworkers allow for the mineral and nonmineral associated 

 proteins to be solubilized (Termine et al. 198 1 ). Five proteins 

 from the mineral compartment of developing human bone 

 have been purified and partially characterized (Fisher et al. 

 1987). 



Of greatest potential interest to the paleopathologist is the 

 preservation of serum-derived proteins in bone. Two serum 

 proteins, albumin and alpha-2-HS. concentrate in bone and 

 make up 13% of the noncollagenous bone matrix proteins 

 extracted from fetal human subperiosteal bone (Robey et al. 

 1988). Smaller amounts of many other serum-derived pro- 

 teins, including transferrin and the immunoglobulins (IgG, 

 IgA, IgM and IgE). can be isolated from modern bone. 



Noncollagenous proteins, osteonectin, albumin. IgG, and 

 transferrin, have been identified at their original molecular 

 weight in several individuals from the Windover archeologi- 

 cal site in Florida. This 7000-year-old site, excavated under 

 the direction of Glen Doran, yielded in excess of 150 human 

 skeletons. The preservation of the mineralized tissue from 

 the Windover site was variable. Generally, however, the neu- 

 tral peat environment provided an anoxic, reducing atmo- 

 sphere that was conducive to protein preservatit)n in these 

 bones. 



This study examines the preservation of noncollagenous 

 proteins — osteonectin, IgG, and albumin — in protein ex- 

 tracts from human skeletal material excavated at the Sully 

 and Mobridge sites in South Dakota. These cemeteries were 

 excavated in the 1920s and the 1950s, and the collections 

 reside in the National Museum of Natural History, Smithso- 

 nian Institution. Associated materials al these sites date the 

 skeletal remains at two to three hundred years of age. These 

 skeletons are a classic museum collection, and provide the 

 opportunity to assess the preservation of protein in bones that 

 have been disinterred for 30 to 50 years. 



Zagreb Paleopathology Symp. 1988 



51 



