Purification, characterization and reconstitution of CMP-N-acetylneuraminate hydroxylase from mouse liver.
CMP-N-acetylneuraminate hydroxylase was isolated from mouse liver high speed supernatant with a yield of 0.4% and an apparent 1000-fold purification. The enzyme is a monomeric protein with a molecular weight of 66 kDa, as determined by gel filtration and SDS-PAGE. The hydroxylase system was reconstituted with Triton X-100-solubilized mouse liver microsomes and purified soluble or microsomal forms of cytochrome b5 reductase and cytochrome b5. The systems were characterized in detail and kinetic parameters for each system were determined.
Biochemisches Institut, Christian-Albrechts-Universität zu Kiel, FRG.
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P
Schneckenburger
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L
Shaw
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R
Schauer
D000818:Animals; D002850:Chromatography, Gel; D003579:Cytochrome Reductases / Q000302:isolation & purification / Q000378:metabolism; D015786:Cytochromes b5 / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006900:Hydroxylation; D007700:Kinetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008862:Microsomes, Liver / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000302:isolation & purification* / Q000378:metabolism
D015786:Cytochromes b5; D006899:Mixed Function Oxygenases; C074383:CMPacetylneuraminate monooxygenase; D003579:Cytochrome Reductases