Partial purification of rat liver microsomal glucose-6-phosphatase on hydroxylapatite.
Glucose-6-phosphatase was effectively solubilized from rat liver-microsomal membrane by the nonionic detergent Renex 690 in the presence of 0.6M sodium chloride. Subsequent separation on hydroxylapatite proved to be a successful and rapid initial step towards the purification of this enzyme. Glucose-6-phosphatase appeared in the colourless void volume with a yield of about 40-50%. The specific activity in the pooled void volume was 3-4 U/mg protein representing an enrichment of 30- to 40-fold. The best final specific activity obtained in an enriched fraction was 6.7 U/mg protein. Analysis of the pooled glucose-6-phosphatase-enriched fraction by SDS electrophoresis revealed 2 dominant protein bands with the apparent molecular mass of 17 and 18.5 kDa and few weak protein bands in the range of 21 to 42 kDa.
Rymsa
B
B
Institut für Physiologische Chemie I, Universität Düsseldorf.
de Groot
H
H
D000818:Animals; D002845:Chromatography / Q000379:methods; D017886:Durapatite; D005952:Glucose-6-Phosphatase / Q000302:isolation & purification* / Q000378:metabolism; D006882:Hydroxyapatites; D007700:Kinetics; D008081:Liposomes; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D008970:Molecular Weight; D010743:Phospholipids / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012995:Solubility
D006882:Hydroxyapatites; D008081:Liposomes; D010743:Phospholipids; D017886:Durapatite; D005952:Glucose-6-Phosphatase