Unusual properties of the prespore-specific enzyme, UDPgalactose:polysaccharide galactosyl transferase, of Dictyostelium discoideum.
UDPgalactose:polysaccharide galactosyl-transferase is the enzyme that is specifically localized in prespore cells of Dictyostelium discoideum and its activity sharply changes in response to differentiation and dedifferentiation. To clarify the nature of this enzyme, we first developed an improved assay method for the enzyme, and by using this method, we partially purified the enzyme through DEAE-sepharose, phenyl-sepharose and ATP-sepharose chromatography. The apparent molecular mass of the enzyme was ca. 200 KDa (by non-denaturing polyacrylamide gel gradient analysis) and the isoelectric point was around pH 7. The enzyme exhibited a hitherto undescribed property, that is the reaction proceeds faster at 0 degrees C than at 21 degrees C, with a smaller K(m) value and an unchanged V(max) value. This low-temperature resistant property of the enzyme is consistent with the previous observation (Maeda 1984, J. Cell Sci. 69, 159-165) that prespore differentiation is favored at low temperatures. The reaction appears to proceed in a double displacement manner. ATP reversibly inhibited the enzyme with a K(i) value of 2 mM, suggesting the possibility that ATP regulates its activity in vivo.
Nakahara
Yasuo
Y
Department of Botany, Faculty of Science, Kyoto University, Kyoto 606-8502, Japan.
Okamoto
Koji
K
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004023:Dictyostelium / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D005700:Galactosyltransferases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D007526:Isoelectric Point; D008970:Molecular Weight; D033761:Spores, Protozoan / Q000201:enzymology; D013696:Temperature
D004791:Enzyme Inhibitors; D000255:Adenosine Triphosphate; D005700:Galactosyltransferases; C023239:lipopolysaccharide galactosyltransferase