title: Anionic phospholipids are determinants of membrane protein topology.
abstract: The orientation of many membrane proteins is determined by the asymmetric distribution of positively charged amino acid residues in cytoplasmic and translocated loops. The positive-inside rule states that loops with large amounts of these residues tend to have cytoplasmic locations. Orientations of constructs derived from the inner membrane protein leader peptidase from Escherichia coli were found to depend on the anionic phospholipid content of the membrane. Lowering the contents of anionic phospholipids facilitated membrane passage of positively charged loops. On the other hand, elevated contents of acidic phospholipids in the membrane rendered translocation more sensitive to positively charged residues. The results demonstrate that anionic lipids are determinants of membrane protein topology and suggest that interactions between negatively charged phospholipids and positively charged amino acid residues contribute to the orientation of membrane proteins.
authors.affiliation: Department of Biochemistry of Membranes, Centre for Biomembranes and Lipid Enzymology, Utrecht University, The Netherlands.
authors.forename: W
authors.initials: W
authors.lastname: van Klompenburg
authors.forename: I
authors.initials: I
authors.lastname: Nilsson
authors.forename: G
authors.initials: G
authors.lastname: von Heijne
authors.forename: B
authors.initials: B
authors.lastname: de Kruijff
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D000838:Anions / Q000737:chemistry; D001386:Azides / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry* / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins; D015151:Immunoblotting; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D008239:Lysine / Q000737:chemistry; D008563:Membrane Lipids / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000737:chemistry* / Q000378:metabolism; D011233:Precipitin Tests; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019810:Sodium Azide; D014357:Trypsin / Q000378:metabolism
chemical_list: D000838:Anions; D001386:Azides; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D008563:Membrane Lipids; D008565:Membrane Proteins; D010743:Phospholipids; D011993:Recombinant Fusion Proteins; C043426:inner membrane protein, E coli; D007544:Isopropyl Thiogalactoside; D019810:Sodium Azide; D000469:Alkaline Phosphatase; D012697:Serine Endopeptidases; D014357:Trypsin; C026515:type I signal peptidase; D008239:Lysine