Solubilization and stabilization of human liver glycoprotein sialyltransferase.
Publication/Presentation Date
9-12-1979
Abstract
Triton X-100 is increasingly effective in solubilizing human liver glycoprotein (asialofetuin) sialytransferase (CMP-N-acetylneuraminate:D-galactosyl-glycoprotien N-acetylneuraminyltransferase, EC 2.4.99.1) activity as its concentration is increased in the homogenizing buffer. At the optimal concentration of 1.5% (v/v), essentially all of the homogenate sialyltransferase activity was solubilized into the supernatant fluid. Higher concentrations of Triton X-100 inhibited sialyltransferase activity. Several kinetic properties of the solubilized asialofetuin-sialyltransferase activity were compared to those of the membrane-bound enzyme(s) (in homogenates made without Triton X-100 or in resuspended pellets). No major difference was apparent, suggesting that solubilization has not significantly altered the properties of sialyltransferase. The solubilized sialyltransferase activity is quite unstable, losing approximately 50% of its activity after one week of storage at 4 degrees C. Various detergents (Zwittergent, sodium taurocholate and sodium deoxycholate) are differentially effective in stabilizing the solubilized activity. Sodium taurocholate (1.5%, w/v) was most effective with no loss in activity after 40 days and minimal loss (14%) after 60 days storage at 4 degrees C. The solubilized sialyltransferase preparation retains full activity after storage in the frozen state (-20 degrees C) for at least 159 days.
Volume
570
Issue
1
First Page
56
Last Page
64
ISSN
0006-3002
Published In/Presented At
Alhadeff, J. A., & Holzinger, R. T. (1979). Solubilization and stabilization of human liver glycoprotein sialyltransferase. Biochimica et biophysica acta, 570(1), 56–64. https://doi.org/10.1016/0005-2744(79)90200-6
Disciplines
Medicine and Health Sciences
PubMedID
39614
Department(s)
Department of Medicine
Document Type
Article