Identification of phospholipase D from cabbage as N-terminally acetylated PLD2.
Recently, the genes of two isoenzymes of phospholipase D from white cabbage (PLD1 and PLD2) with molecular masses of 91.7 and 91.9 kDa, respectively, have been sequenced and expressed in Escherichia coli [Schäffner, I., Rücknagel, K.-P., Mansfeld, J., and Ulbrich-Hofmann, R. (2002). Eur. J. Lipid Sci. Technol. 104: 79-87]. Both enzymes are highly homologous (91% identity) and behave very similarly. Phospholipase D purified from white cabbage leaves (PLDcab) is compared with the two recombinant enzymes in sodium dodecylsulfate and native polyacrylamide gel electrophoresis, isoelectric focusing, N-terminal sequencing, and mass spectrometry after tryptic digestion. As a result, PLDcab clearly can be assigned to PLD2. In contrast to recombinant PLD2, however, PLDcab is N-terminally acetylated.
Schöps
Regina
R
Martin-Luther University Halle-Wittenberg, Department of Biochemistry/Biotechnology, Kurt-Mothes-Str. 3, D-06120 Halle, Germany.
Schierhorn
Angelika
A
Schäffner
Ines
I
Mansfeld
Johanna
J
Ulbrich-Hofmann
Renate
R
D000107:Acetylation; D001937:Brassica / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D010739:Phospholipase D / Q000737:chemistry* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D010739:Phospholipase D