Details

Autor(en) / Beteiligte

• Cathala, G
• Brunel, C

Titel

Bovine kidney alkaline phosphatase. Purification, subunit structure, and metalloenzyme properties

Ist Teil von

• The Journal of biological chemistry, 1975-08, Vol.250 (15), p.6040-6045

Ort / Verlag

United States: American Society for Biochemistry and Molecular Biology

Erscheinungsjahr

1975

Quelle

MEDLINE

Beschreibungen/Notizen

• Kidney alkaline phosphatase was purified to homogeneity. It is a glycoprotein of about 172,000 molecular weight. Analyses of the subunit structure by sedimentation equilibrium in 6 M guanidine hydrochloride and by gel electrophoresis in sodium dodecyl sulfate indicate that the alkaline phosphatase is a dimer comprising two very similar or identical subunits of about 87,000 molecular weight. The native enzyme contains 4.5 +/- 0.2 g atoms of zinc per mol of protein. Reconstitution experiments from the apophosphatase show that binding of 4 Zn2+ per mol of dimer is essential for full activity. The kinetic data of Zn2+ binding to the apoprotein require at least a two-step mechanism, in which one of the steps corresponds to a conformational change within the enzyme. This paper also presents data concerning amino acid composition, sugar content, enzyme stability, absorbance index, and sedimentation velocity.