Immuno-identification of two non-amelogenin proteins of developing bovine enamel isolated by affinity chromatography. Further proof that tooth "enamelins" are mainly serum proteins.
Affinity chromatography of "Enamelin Extracts" of developing bovine molar enamel on CNBr activated Sepharose 4B to which polyclonal antibodies of whole bovine serum and fetuin were cross-linked, revealed that at most, only 1-2% of the proteins in the extracts were not bound to the columns. The approximately 98% or more of the proteins in such extracts were bound to the resin and were eluted in the position of the serum proteins and fetuin. The small amount of protein which was not bound to the affinity column and which was eluted very early, was subjected to SDS-PAGE and immunostained with polyclonal antibodies to two non-amelogenin proteins isolated and identified previously (approximately 26kDa and 22kDa). The antibody to the approximately 50kDa protein immunostained strongly with only one protein band of approximately 26kDa. The antibody to the approximately 22kDa band reacted strongly with one protein band of approximately 22kDa, weakly with an approximately 100kDa and very weakly with several lower molecular weight bands, suggesting either aggregation and degredation of the 22kDa component, or more likely that the approximately 22kDa and lower molecular weight proteins are all derived from the approximately 100kDa component. There was no immuno-crossreactivity of the 22kDa and 26kDa antibodies and none of the components eluted in the early first fraction reacted with polyclonal antibodies to amelogenins or to amelogenin peptides.(ABSTRACT TRUNCATED AT 250 WORDS)
Strawich
E
E
Department of Orthopedic Surgery, Harvard Medical School, Children's Hospital, Boston, MA 02115.
Seyer
J
J
Glimcher
M J
MJ
D053523:Amelogenin; D000818:Animals; D001798:Blood Proteins / Q000502:physiology*; D002417:Cattle; D002846:Chromatography, Affinity; D003743:Dental Enamel / Q000737:chemistry* / Q000254:growth & development*; D003746:Dental Enamel Proteins / Q000302:isolation & purification* / Q000502:physiology*; D007158:Immunologic Techniques
D053523:Amelogenin; D001798:Blood Proteins; D003746:Dental Enamel Proteins