Rat alkaline phosphatase. II. Structural similarities between the osteosarcoma, bone, kidney, and placenta isoenzymes.
A mouse monoclonal antibody raised against rat osteosarcoma alkaline phosphatase (AP) was covalently coupled to protein A-Sepharose and used to purify this enzyme from preparations of rat osteosarcoma, calvaria, kidney, and placenta in a single-step procedure. The tissue-specific isoenzymes purified in this manner showed identity in the immunodiffusion reaction with a polyclonal anti-AP antibody, but differed in apparent molecular weight and degree of polydispersity on sodium dodecyl sulfate-polyacrylamide gels. Treatment with N-glycanase abolished these differences, yielding proteins with an apparent molecular weight of 52,000 Da and identical V8 protease digestion patterns. Alkaline phosphatase from these tissues showed no significant difference in amino acid composition and identity in the first 20 N-terminal amino acids. These findings provide structural evidence which supports the hypothesis that the tissue-specific alkaline phosphatase isoenzymes share a common protein sequence subject to different glycosylation pattern.
Nair
B C
BC
Johnson
D E
DE
Majeska
R J
RJ
Rodkey
J A
JA
Bennett
C D
CD
Rodan
G A
GA
D000469:Alkaline Phosphatase / Q000276:immunology / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001842:Bone and Bones / Q000201:enzymology; D005260:Female; D007120:Immunochemistry; D007527:Isoenzymes / Q000276:immunology / Q000302:isolation & purification*; D007668:Kidney / Q000201:enzymology; D008970:Molecular Weight; D012516:Osteosarcoma / Q000201:enzymology; D010920:Placenta / Q000201:enzymology; D011247:Pregnancy; D051381:Rats; D011912:Rats, Inbred ACI
D000596:Amino Acids; D007527:Isoenzymes; D000469:Alkaline Phosphatase