Purification and partial amino acid sequencing of rat bone tumor (UMR106) alkaline phosphatase.
Cultured rat osteosarcoma (UMR106) alkaline phosphatase was purified to apparent homogeneity by sequential application of polyclonal antibody affinity, DEAE-cellulose, and Sepharose CL-6B chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the enzyme preparation treated with sodium dodecyl sulfate and mercaptoethanol showed the presence of a dominant band (using silver staining) corresponding to a molecular weight of 80,000. The amino acid composition was similar to those of various alkaline phosphatases. The N-terminal amino acid sequence was determined as follows: Phe-Val-Pro-Glu-Lys-Glu-Lys- Asp-Pro-Ser-Tyr-Trp-Arg-Gln-Gln-Ala-Gln-Glu-Thr-Leu- Lys-Asn-Ala-Leu-Lys-?-Gln-Lys-?-Asn-Val-Asn-Ala-Lys.
Hsu
H H
HH
Rouse
J
J
Hamilton
J
J
Anderson
H C
HC
D000469:Alkaline Phosphatase / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001859:Bone Neoplasms / Q000201:enzymology*; D002845:Chromatography / Q000379:methods; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D007158:Immunologic Techniques; D012516:Osteosarcoma / Q000201:enzymology*; D051381:Rats; D012513:Sarcoma, Experimental / Q000201:enzymology*
D000596:Amino Acids; D000469:Alkaline Phosphatase