Nonspecific lipid transfer protein in castor bean cotyledon cells: subcellular localization and a possible role in lipid metabolism.
The subcellular localization and several biochemical activities of nonspecific lipid transfer protein (nsLTP) were investigated. A section of a castor bean cotyledon cell was labeled with anti-nsLTP serum followed by protein A-gold. Gold particles were more abundant in the glyoxysome matrix and the vessel cell wall than in other areas. Cell fractionation analysis of 6-day-old castor bean cotyledons by sucrose density gradient centrifugation demonstrated that 13% of nsLTP was distributed in the glyoxysomal fraction, identified on the basis of catalase as a marker, and 87% in the soluble fraction near the top of the gradient. The location of castor bean nsLTP in glyoxysomes was further confirmed by in vitro import experiments. The synthesized precursor of nsLTP (pro-nsLTP-C) was incorporated into intact castor bean glyoxysomes and processed to the mature form after import into the glyoxysomes, but it was not imported into canine pancreatic microsomes. Castor bean nsLTP-A was found to possess the ability to bind oleic acid and oleoyl-CoA by means of a method involving Lipidex 1000. The dissociation constants (Kd) for oleic acid and oleoyl-CoA binding to nsLTP-A were 4.8 and 5.0 microM, respectively. The saturated binding capacities (Bmax) for oleic acid and oleoyl-CoA per mol of nsLTP-A were 1.1 and 1.2 mol, respectively. When acyl-CoA oxidase activity was assayed in the glyoxysomal fraction, marked enhancement of the activity was observed in the presence of nsLTP. These results suggest the possibility that nsLTP regulates fatty acid beta-oxidation through the enhancement of acyl-CoA oxidase activity in glyoxysomes. The occurrence of castor bean nsLTP in the vessel wall was discussed.
Tsuboi
S
S
Department of Biology, College of Arts and Sciences, University of Tokyo.
Osafune
T
T
Tsugeki
R
R
Nishimura
M
M
Yamada
M
M
D000214:Acyl Coenzyme A / Q000378:metabolism; D042962:Acyl-CoA Oxidase; D002352:Carrier Proteins / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D002367:Ricinus communis / Q000166:cytology / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism; D050356:Lipid Metabolism; D010088:Oxidoreductases / Q000187:drug effects / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism* / Q000494:pharmacology; D010947:Plants, Toxic; D013347:Subcellular Fractions / Q000378:metabolism
D000214:Acyl Coenzyme A; D002352:Carrier Proteins; D005227:Fatty Acids; D010940:Plant Proteins; C072087:sterol carrier proteins; D010088:Oxidoreductases; D042962:Acyl-CoA Oxidase