Characterization and purification of acid phosphatase from ancient human bone.
In this research, acid phosphatase was purified and characterized from approximately 3000-year-old human bones from archeological excavations. Using anion exchange chromatography, two isoenzymes, TrACP and TsACP, were isolated from the bone. TrACP and TsACP were eluted separately, with a concentration gradient, from a CM-sepharose column. The resulting TrACP was further purified on a cellulose phosphate column. The activity was determined by using pNPP as substrate. Additionally, protein was determined by the Bradford and Coomassie Brilliant Blue method. The optimum pHs of TsACP and TrACP were 6 and 5, respectively. The optimum temperatures were 0 and 10 degrees C, respectively. Molecular weights were measured by gel filtration chromatography. The isoenzyme purity was checked with SDS-PAGE. Finally, the effects of sodium molybdate and tartrate on isoenzyme activity were determined.
Demir
Yaşar
Y
Department of Chemistry, Atatürk University, Erzurum, Turkey.
Yildirim
Safinur
S
Alayli
Azize
A
Demir
Nazan
N
D000135:Acid Phosphatase / Q000737:chemistry / Q000266:history / Q000302:isolation & purification*; D001842:Bone and Bones / Q000201:enzymology*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D049690:History, Ancient; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification; D008970:Molecular Weight; D000071681:Tartrate-Resistant Acid Phosphatase; D013696:Temperature
D007527:Isoenzymes; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase