The distribution of soluble, mineral-bound, and matrix-bound proteins in osteoporotic and normal bones.
Very little is known about the composition of noncollagenous protein (NCP) in osteoporotic bone. We have obtained osteoporotic (OP) cancellous bone samples from 2-4 cm below the surface of human femoral heads and compared the protein profiles to age-matched (old controls, OC), as well as young autopsy controls (YC). The proteins were analyzed using a serial extraction method which liberates proteins from distinct pools; first, proteins were extracted in 4.0 M GuHCl (G1), followed by the extraction of mineral-bound protein with 0.5 M EDTA (E), and finally the protein previously masked by the mineral was released by a second 4.0 M GuHCl extraction (G2). The tissue residue was digested with bacterial collagenase liberating proteins bound to the collagenous matrix. Uniform bone preparations of finely ground bone powder were used for our study. The protein content of the four groups was determined together with the protein distribution using amino acid analysis and gel-electrophoresis. Our data indicate that the noncollagenous protein content is considerably reduced in osteoporotic bone when compared with age-matched or young controls, whereas the collagen content is unaffected. When bone proteins were investigated using gel-electrophoresis we found a similar distribution of proteins in the G1 and E fractions of all three groups, but the osteoporotic bone samples, using densitometric analysis of the gels, showed quantitative differences in a number of the proteins. The greatest differences were observed in the G2 extract, where the NCP of osteoporotic bone was limited to a few low-molecular-weight bands when compared with the OCs or YCs.(ABSTRACT TRUNCATED AT 250 WORDS)
Grynpas
M D
MD
Department of Pathology, University of Toronto, Ontario, Canada.
Tupy
J H
JH
Sodek
J
J
D015502:Absorptiometry, Photon; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000596:Amino Acids / Q000378:metabolism; D000704:Analysis of Variance; D015153:Blotting, Western; D015519:Bone Density / Q000502:physiology*; D002851:Chromatography, High Pressure Liquid; D003094:Collagen / Q000378:metabolism; D017364:Collagenases / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005260:Female; D005265:Femoral Neck Fractures / Q000601:surgery; D005270:Femur Head / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D008297:Male; D008875:Middle Aged; D008970:Molecular Weight; D010024:Osteoporosis / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012995:Solubility
D000596:Amino Acids; D016326:Extracellular Matrix Proteins; D011506:Proteins; D003094:Collagen; D017364:Collagenases