Purification and partial amino acid sequencing of alkaline phosphatase from rachitic rat epiphyseal cartilage.
1. Alkaline phosphatase of rachitic epiphyseal cartilage was purified to apparent homogeneity by sequential application of monoclonal affinity, DEAE-cellulose, and Sepharose CL-6B chromatography. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of the enzyme showed the presence of a dominant band corresponding to a molecular weight of 80,000. 2. The N-terminal amino acid sequence was determined as follows: Phe-Val-Pro-Glu-Lys5-Glu-Lys-Asp-Pro-Ser10-Tyr-Trp-Arg-Gln-+ ++Gln15-Ala-Gln-Glu- Thr-Leu20-Lys-Asn-Ala-Leu-Lys25-Leu-Gln-Lys-?-Asn-Val-Asn-?- Ala-Lys35-?-Ile-?- Met-Phe40-Leu-(Gly?)-Asp-(Ala/Gly?)-Met45-?-Val-?- (Val/Gly?).
Hsu
H H
HH
Department of Pathology and Oncology, University of Kansas Medical Center, Kansas City 66103.
Rouse
J
J
Hamilton
J
J
Anderson
H C
HC
D000469:Alkaline Phosphatase / Q000276:immunology / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000302:isolation & purification; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000915:Antibody Affinity; D002356:Cartilage / Q000201:enzymology*; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D051381:Rats; D012279:Rickets / Q000201:enzymology*
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000469:Alkaline Phosphatase