title: Fatty acid acylated peroxidase as a model for the study of interactions of hydrophobically-modified proteins with mammalian cells.
abstract: Artificial fatty acylation of proteins has attracted significant attention during the last decade as a method for modification of protein specificity and efficacy of action on mammalian cells (A. V. Kabanov and V. Yu. Alakhov (1994) J. Contr. Release 28, 15-35). Horse radish peroxidase (HRP) is used in this work to study the interaction of a fatty acylated protein with various mammalian cells. The HRP is modified with the chloranhydride of the stearic acid in the reversed micelles of sodium bis-(2-ethylhexyl)sulfosuccinate (Aerosol OT) in octane, a convenient protocol allowing production of protein molecules with a controlled, low modification degree (A.V. Kabanov et al. (1987) Ann. N. Y. Acad. Sci. 501, 63-66). The influence of the hydrophobic group on the binding and internalization of HRP in MDCK, P3-X63-Ag8, CHO, and HepG2 cells is examined. The major results are as follows: (i) the fatty acylation of a protein significantly enhances its binding to all tested mammalian cell lines, with a line-specific efficiency; (ii) the binding efficiency can be modified by changing growth conditions in a defined medium; (iii) along with the enhancement of protein adsorption on the plasma membrane, fatty acylation increases internalization of the protein during incubations at 37 degrees C; (iv) internalized protein was observed in endocytic vesicles; no evidence was obtained for a cytoplasmic distribution. These results are discussed in connection with previously observed effects of the fatty acylated proteins on cell activity.
authors.affiliation: Laboratory of Biopolymer Chemistry, Russian Research Center of Molecular Diagnostics and Therapy (RCMDT), Moscow, Russia.
authors.forename: V I
authors.initials: VI
authors.lastname: Slepnev
authors.forename: L
authors.initials: L
authors.lastname: Phalente
authors.forename: H
authors.initials: H
authors.lastname: Labrousse
authors.forename: N S
authors.initials: NS
authors.lastname: Melik-Nubarov
authors.forename: V
authors.initials: V
authors.lastname: Mayau
authors.forename: B
authors.initials: B
authors.lastname: Goud
authors.forename: G
authors.initials: G
authors.lastname: Buttin
authors.forename: A V
authors.initials: AV
authors.lastname: Kabanov
mesh_terms: D000215:Acylation; D000327:Adsorption; D000818:Animals; D001693:Biological Transport, Active; D016466:CHO Cells; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D004285:Dogs; D005227:Fatty Acids / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D051379:Mice; D008823:Micelles; D008954:Models, Biological; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D005227:Fatty Acids; D008823:Micelles; D011506:Proteins; D006735:Horseradish Peroxidase