A phospholipase A2 is transiently synthesized during seed germination and localized to lipid bodies.
A patatin-like protein is present in the storage tissue of cucumber seedlings during the stage of fat mobilization. The cucumber protein is a homologue of a glycoprotein which in potatoes accounts for most of the total protein content of tubers. Following preparation of a cucumber cDNA library representing the developmental stage of cotyledons of 1 day old germinating seeds we isolated and characterized a clone encoding a patatin-like protein. Antibodies raised against the protein expressed in bacteria were used for immunodetection in subcellular fractions of cucumber seedlings. It was shown that the patatin-like protein was virtually exclusively confined to lipid bodies. The protein expressed in bacteria was characterized in vitro by its esterase activity acting on monoacylglycerols and phospholipids. Detailed analysis using various forms of phosphatidyl choline as substrates demonstrated that the patatin-like protein is a phospholipase A2 acting on palmitoyl, linoleoyl and hydroperoxidized linoleoyl groups equally well. Studying the temporal and tissue-specific expression of patatin-like protein mRNA we showed its appearance exclusively during fat catabolism. As maximal amounts of the protein were found at an early stage of mobilization and confined to lipid bodies, we propose that the patatin-like hydrolase is involved in lipid body mobilization.
May
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C
Fachbereich Chemie, Philipps-Universität, Marburg, Germany. hyola@ps1515.chemie.uni-marburg.de
Preisig-Müller
R
R
Höhne
M
M
Gnau
P
P
Kindl
H
H
D000595:Amino Acid Sequence; D002265:Carboxylic Ester Hydrolases; D018548:Cotyledon / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D015723:Gene Library; D018525:Germination; D050356:Lipid Metabolism; D008969:Molecular Sequence Data; D010741:Phospholipases A / Q000096:biosynthesis* / Q000235:genetics; D054467:Phospholipases A2; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D012639:Seeds; D014675:Vegetables
D010940:Plant Proteins; D012333:RNA, Messenger; C043222:patatin protein, Solanum tuberosum; D002265:Carboxylic Ester Hydrolases; D010741:Phospholipases A; D054467:Phospholipases A2