The precursor form of the rat liver non-specific lipid-transfer protein, expressed in Escherichia coli, has lipid transfer activity.
The cDNA encoding the precursor form of non-specific lipid-transfer protein (pre-nsL-TP) from rat liver was cloned into the expression vector pET3d. The resulting plasmid was transformed to the Escherichia coli strain BL21(DE3). After induction of the bacteria with isopropyl-beta-D-thiogalactopyranoside (IPTG) pre-nsL-TP was purified from the bacterial lysate by anion exchange chromatography followed by gelfiltration. From 11 of culture, 6-7 mg of pre-nsL-TP was obtained, equal to approximately 7% of the cytoplasmic protein. By use of a fluorescence lipid transfer assay, pre-nsL-TP was found to have lipid transfer activity identical to mature nsL-TP.
Ossendorp
B C
BC
Centre for Biomembranes and Lipid Enzymology, State University of Utrecht, The Netherlands.
Geijtenbeek
T B
TB
Wirtz
K W
KW
D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008099:Liver / Q000737:chemistry; D008969:Molecular Sequence Data; D010713:Phosphatidylcholines / Q000378:metabolism*; D010940:Plant Proteins; D011498:Protein Precursors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
D002352:Carrier Proteins; D010713:Phosphatidylcholines; D010940:Plant Proteins; D011498:Protein Precursors; D011994:Recombinant Proteins; C072087:sterol carrier proteins; C021248:1-palmitoyl-2-pyrenedecanoylphosphatidylcholine