A 138-kDa glycoprotein from Dictyostelium membranes with folate deaminase and folate binding activity.
A 138-kDa glycoprotein comprising folate deaminase activity was purified to apparent homogeneity from membranes of Dictyostelium discoideum. Deaminase activity could be effectively inhibited by p-chloromercuriphenylsulfonate. This treatment protected folate from deamination and thus allowed investigation of folate binding to deaminase fractions. Two types of folate binding sites, differing in affinity and specificity, were detected on the folate deaminase glycoprotein. One type displays high affinity and binds folate stronger than N10-methylfolate. This binding site appears to be identical with the catalytic site of folate deaminase. The other type of binding site shows lower affinity but prefers N10-methylfolate relative to folate. A similar preference for N10-methylfolate was observed in chemotaxis tests pointing to the possibility that the second type of binding site is involved in chemotactic perception of folate compounds. Folate perception and deamination could thus be performed by activities residing on the same polypeptide.
Greiner
R A
RA
Fakultät für Biologie, Universität Konstanz, Federal Republic of Germany.
Jacobs-Krahnen
D
D
Mutzel
R
R
Malchow
D
D
Wurster
B
B
D000619:Aminohydrolases / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D004023:Dictyostelium / Q000201:enzymology / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D058973:Folate Receptors, GPI-Anchored; D005492:Folic Acid / Q000378:metabolism; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D015800:Protozoan Proteins; D011956:Receptors, Cell Surface
D002352:Carrier Proteins; D058973:Folate Receptors, GPI-Anchored; D006023:Glycoproteins; D015800:Protozoan Proteins; D011956:Receptors, Cell Surface; D005492:Folic Acid; D000619:Aminohydrolases; C020144:folic acid deaminase, Dictyostelium discoideum