Activity increase after extraction of alkaline phosphatase from human osteoblastic membranes by nonionic detergents: influence of age and sex.
The solubilization of alkaline phosphatase (AP) from osteoblastic cell membranes obtained from human primary bone cell cultures was studied according to the age and sex of the donors (17 females, 11 males; age range: 2-77 years). Cell membranes were treated by non-ionic (n-octyl beta-D-glucopyranoside, OG), ionic or zwitterionic detergents, then centrifuged. When OG was used almost all the AP was solubilized. AP activity in supernatant of solubilization was compared to the activity of the suspension before centrifugation. The activity ratio (AR) increased in function of age for subjects between 65 and 74. Neither total nor specific AP activities were influenced by age or sex. Electrophoresis studies showed that the AP released was a GPI (glycosyl phosphatidylinositol)-anchored protein, amphipathic form, with 140 kDa as apparent molecular mass. The activity change of AP in the presence of OG may result from age-related modifications either in the AP structure or in the constituents of the plasma membranes (proteins or phospholipids).
Bourrat
C
C
Université Claude Bernard Lyon I, UPRESA CNRS 5013, 43 bd du 11 Novembre 1918, 69622 Villeurbanne Cedex, France.
Radisson
J
J
Chavassieux
P
P
Azzar
G
G
Roux
B
B
Meunier
P J
PJ
D000293:Adolescent; D000328:Adult; D000367:Age Factors; D000368:Aged; D000469:Alkaline Phosphatase / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003902:Detergents / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005960:Glucosides / Q000494:pharmacology*; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D006801:Humans; D008297:Male; D008875:Middle Aged; D008970:Molecular Weight; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D012737:Sex Factors; D012995:Solubility
D003902:Detergents; D005960:Glucosides; D017261:Glycosylphosphatidylinositols; C018619:octyl-beta-D-glucoside; D000469:Alkaline Phosphatase