title: Expression and purification of recombinant human apolipoprotein A-II in Pichia pastoris.
abstract: Apolipoprotein A-II (ApoA-II) is the second most abundant protein constituent of high-density lipoprotein (HDL). The physiologic role of ApoA-II is poorly defined. ApoA-II may inhibit lecithin:cholesterol acyltransferase and cholesteryl-ester-transfer protein activities, but may increase the hepatic lipase activity. ApoA-II may also inhibit the hepatic cholesteryl uptake from HDL probably through the scavenger receptor class B type I depending pathway. Interpretation of data from transgenic and knockout mice of genes involved in lipoprotein metabolism has been often complicated as clinical implications because of species difference. So it is important to obtain human ApoA-II for further studies about its functions. In our studies, Pichia pastoris expression system was first used to express a high-level secreted recombinant human ApoA-II (rhApoA-II). We have cloned the cDNA encoding human ApoA-II and achieved its high-level secreting expression with a yield of 65 mg/L of yeast culture and the purification process was effective and easy to handle. The purified rhApoA-II can be used to further study its biological activities.
authors.affiliation: 1 Department of Regenerative Medicine, College of Pharmacy, Jilin University , Changchun, China .
authors.forename: Manman
authors.initials: M
authors.lastname: Su
authors.forename: Yitian
authors.initials: Y
authors.lastname: Qi
authors.forename: Mingxing
authors.initials: M
authors.lastname: Wang
authors.forename: Weiqin
authors.initials: W
authors.lastname: Chang
authors.forename: Shuang
authors.initials: S
authors.lastname: Peng
authors.forename: Tianmin
authors.initials: T
authors.lastname: Xu
authors.forename: Dingding
authors.initials: D
authors.lastname: Wang
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D016633:Apolipoprotein A-II / Q000096:biosynthesis*; D015153:Blotting, Western; D003001:Cloning, Molecular; D003470:Culture Media; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005285:Fermentation; D005822:Genetic Vectors; D006801:Humans; D007202:Indicators and Reagents; D008081:Liposomes / Q000737:chemistry; D000432:Methanol / Q000378:metabolism; D010843:Pichia / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D016633:Apolipoprotein A-II; D003470:Culture Media; D018076:DNA, Complementary; D007202:Indicators and Reagents; D008081:Liposomes; D011994:Recombinant Proteins; D000429:Alcohol Oxidoreductases; D000432:Methanol