title: A hydrophobic patch in a charged alpha-helix is sufficient to target proteins to dense core secretory granules.
abstract: Many endocrine and neuroendocrine cells contain specialized secretory organelles called dense core secretory granules. These organelles are the repository of proteins and peptides that are secreted in a regulated manner when the cell receives a physiological stimulus. The targeting of proteins to these secretory granules is crucial for the generation of certain peptide hormones, including insulin and ACTH. Although previous work has demonstrated that proteins destined to a variety of cellular locations, including secretory granules, contain targeting sequences, no single consensus sequence for secretory granule-sorting signals has emerged. We have shown previously that alpha-helical domains in the C-terminal tail of the prohormone convertase PC1/3 play an important role in the ability of this region of the protein to direct secretory granule targeting (Jutras, I. Seidah, N. G., and Reudelhuber, T. L. (2000) J. Biol. Chem. 275, 40337-40343). In this study, we show that a variety of alpha-helical domains are capable of directing a heterologous secretory protein to granules. By testing a series of synthetic alpha-helices, we also demonstrate that the presence of charged (either positive or negative) amino acids spatially segregated from a hydrophobic patch in the alpha-helices of secretory proteins likely plays a critical role in the ability of these structures to direct secretory granule sorting.
authors.affiliation: Laboratory of Molecular Biochemistry of Hypertension, Clinical Research Institute of Montreal, Montreal, Quebec H2W 1R7, Canada.
authors.forename: Jimmy D
authors.initials: JD
authors.lastname: Dikeakos
authors.forename: Marie-Josée
authors.initials: MJ
authors.lastname: Lacombe
authors.forename: Chantal
authors.initials: C
authors.lastname: Mercure
authors.forename: Matei
authors.initials: M
authors.lastname: Mireuta
authors.forename: Timothy L
authors.initials: TL
authors.lastname: Reudelhuber
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D006728:Hormones / Q000737:chemistry / Q000378:metabolism; D057927:Hydrophobic and Hydrophilic Interactions; D051379:Mice; D008969:Molecular Sequence Data; D009490:Neurosecretory Systems / Q000166:cytology / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000502:physiology*; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D022142:Secretory Vesicles / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D006728:Hormones; D021382:Protein Sorting Signals; D011506:Proteins; D011993:Recombinant Fusion Proteins