title: Purification and characterization of human nucleolar phosphoprotein 140 expressed in Escherichia coli.
abstract: Human nucleolar phosphoprotein 140, hNopp140, is one of the most highly phosphorylated mammalian proteins, which is involved in the biogenesis of nucleolus. It regulates the transcription of rDNA and has a tendency to bind to doxorubicin, which is widely used as an anti-cancer drug. The biochemical and biophysical property of hNopp140 has not been reported due to the fact that it is rather difficult to obtain protein in large enough quantity. In this paper, we report the cloning and overexpression of the soluble form of hNopp140 in Escherichia coli. The protein was purified to more than 90% homogeneity using hydroxyapatite and ion exchange chromatography. The purified protein can be extensively phosphorylated by casein kinase II and oligomerized into an insoluble aggregate in the presence of magnesium, carbonate, and fluoride ions.
authors.affiliation: Division of Life Sciences, Korea Institute of Science and Technology, 39-1, Hwawolkok-dong, Songbuk-ku, 136-791 Seoul, South Korea.
authors.forename: Yun-Kyeong
authors.initials: YK
authors.lastname: Kim
authors.forename: Youngnam
authors.initials: Y
authors.lastname: Jin
authors.forename: Krishna M
authors.initials: KM
authors.lastname: Vukoti
authors.forename: Joon Kyu
authors.initials: JK
authors.lastname: Park
authors.forename: Eunice E
authors.initials: EE
authors.lastname: Kim
authors.forename: Kong-Joo
authors.initials: KJ
authors.lastname: Lee
authors.forename: Yeon Gyu
authors.initials: YG
authors.lastname: Yu
mesh_terms: D000818:Animals; D001483:Base Sequence; D047390:Casein Kinase II; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005459:Fluorides / Q000494:pharmacology; D006801:Humans; D008274:Magnesium / Q000502:physiology; D009687:Nuclear Proteins / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D010766:Phosphorylation; D015202:Protein Engineering; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; C075564:NOLC1 protein, human; D009687:Nuclear Proteins; D010750:Phosphoproteins; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D008274:Magnesium; D005459:Fluorides