title: Expression and characterization of Cryptococcus neoformans recombinant App1.
abstract: We characterized Cryptococcus neoformans recombinant antiphagocytic protein 1 (rApp1) by SDS-PAGE, gel filtration chromatography, circular dichroism, and fluorescence spectroscopy. rApp1 produced by C. neoformans var. grubii contains an odd number of cysteines; therefore, it has the ability to form intermolecular disulfide bridges which can lead to the formation of amyloid fibrils in the absence of β-mercaptoethanol or DTT in vitro. Alternate approaches to over-expression of rApp1 in the Lepidopteran High Five(™) Insect cell line using pIZ/V5-His and in lentivirus were explored and are described. Finally, localization of App1 in vivo was examined in the presence and absence of the capsule.
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC 29425, USA.
authors.forename: Asfia
authors.initials: A
authors.lastname: Qureshi
authors.forename: Virginia
authors.initials: V
authors.lastname: Williams
authors.forename: Maurizio
authors.initials: M
authors.lastname: Del Poeta
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D002460:Cell Line; D002850:Chromatography, Gel; D002942:Circular Dichroism; D004220:Disulfides; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D005822:Genetic Vectors; D007313:Insecta; D016086:Lentivirus / Q000235:genetics; D008968:Molecular Conformation; D008970:Molecular Weight; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013050:Spectrometry, Fluorescence
chemical_list: C535050:App1 protein, Cryptococcus neoformans; D002352:Carrier Proteins; D004220:Disulfides; D011994:Recombinant Proteins