title: Metal ion requirement of bifunctional UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase from rat liver.
abstract: The metal ion requirement for both enzymatic activities of the bifunctional UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (E.C. 5.1.3.14/2.7.1.60), the key enzyme of N-acetylneuraminic acid biosynthesis in rat liver, was investigated. UDP-N-acetylglucosamine 2-epimerase was active in imidazole/HCl buffer in the complete absence of any metal ion. 200 mM Na+, K+, Rb+ and Cs+ activated enzyme activity up to five-fold, whereas lower concentrations of these monovalent metal ions showed only a small effect on UDP-N-acetylglucosamine 2-epimerase activity. In sodium phosphate buffer the enzyme activity was increased by 0.5 mM Mg2+, Sr2+, Ba2+ and Mn2+, while in the presence of 200 mM NaCl UDP-N-acetylglucosamine 2-epimerase activity showed a stronger activation by these divalent metal ions. In imidazole/HCl buffer, UDP-N-acetylglucosamine 2-epimerase activity was partially inhibited by 0.5 mM Be2+, Mg2+, Ba2+, Mn2+, Sn2+ and Fe2+, and completely inhibited by 0.5 mM Zn2+ and Cd2+. Divalent metal ions were essential for N-acetylmannosamine kinase activity, the most effective being Mg2+, followed by Mn2+ and Co2+. The optimal concentration of these metal ions was 3 mM. Less effective were Ni2+ and Cd2+, whereas Ca2+, Ba2+, Cu2+, Fe2+ and Zn2+ showed no effect on enzyme activity.
authors.affiliation: Institut für Molekularbiologie und Biochemie, Freie Universität Berlin, Dahlem, Germany. hinderli@zedat.fu-berlin.de
authors.forename: S
authors.initials: S
authors.lastname: Hinderlich
authors.forename: A
authors.initials: A
authors.lastname: Sonnenschein
authors.forename: W
authors.initials: W
authors.lastname: Reutter
mesh_terms: D000818:Animals; D002238:Carbohydrate Epimerases / Q000737:chemistry / Q000378:metabolism; D029968:Escherichia coli Proteins; D008099:Liver / Q000201:enzymology*; D008670:Metals / Q000737:chemistry / Q000378:metabolism*; D051381:Rats
chemical_list: D029968:Escherichia coli Proteins; D008670:Metals; D002238:Carbohydrate Epimerases; C100033:UDP acetylglucosamine-2-epimerase; C083982:wecB protein, E coli