title: Apolipoprotein A-I amyloidogenic variant L174S, expressed and isolated from stably transfected mammalian cells, is associated with fatty acids.
abstract: Sixteen variants of apolipoprotein A-I (ApoA-I) are associated with hereditary systemic amyloidoses, characterized by amyloid deposition in peripheral organs of patients. As these are heterozygous for the amyloidogenic variants, their isolation from plasma is impracticable and recombinant expression systems are needed. Here we report the expression of recombinant ApoA-I amyloidogenic variant Leu174 with Ser (L174S) in stably transfected Chinese hamster ovary-K1 cells. ApoA-I variant L174S was found to be efficiently secreted in the culture medium, from which it was isolated following a one-step purification procedure. Mass spectrometry analyses allowed the qualitative and quantitative definition of the amyloidogenic variant lipid content, which was found to consist of two saturated and two monounsaturated fatty acids. Interestingly, the same lipid species were found to be associated with the wild-type ApoA-I, expressed and isolated using the same cell system, with lower values of the lipid to protein molar ratios with respect to the amyloidogenic variant. A possible role of fatty acids in trafficking and secretion of apolipoproteins may be hypothesized.
authors.affiliation: Department of Structural and Functional Biology, University of Naples Federico II, School of Biotechnological Sciences, Naples, Italy.
authors.forename: Daria Maria
authors.initials: DM
authors.lastname: Monti
authors.forename: Sonia
authors.initials: S
authors.lastname: Di Gaetano
authors.forename: Rita
authors.initials: R
authors.lastname: Del Giudice
authors.forename: Chiara
authors.initials: C
authors.lastname: Giangrande
authors.forename: Angela
authors.initials: A
authors.lastname: Amoresano
authors.forename: Maria
authors.initials: M
authors.lastname: Monti
authors.forename: Angela
authors.initials: A
authors.lastname: Arciello
authors.forename: Renata
authors.initials: R
authors.lastname: Piccoli
mesh_terms: D058227:Amyloidogenic Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D000818:Animals; D016632:Apolipoprotein A-I / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D016466:CHO Cells; D018929:Cell Culture Techniques; D002853:Chromatography, Liquid; D006224:Cricetinae; D017077:Culture Media, Conditioned / Q000737:chemistry; D016895:Culture Media, Serum-Free; D005227:Fatty Acids / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D020125:Mutation, Missense; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D058227:Amyloidogenic Proteins; D016632:Apolipoprotein A-I; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D005227:Fatty Acids; D011994:Recombinant Proteins