title: Identification of Human N-Myristoylated Proteins from Human Complementary DNA Resources by Cell-Free and Cellular Metabolic Labeling Analyses.
abstract: To identify physiologically important human N-myristoylated proteins, 90 cDNA clones predicted to encode human N-myristoylated proteins were selected from a human cDNA resource (4,369 Kazusa ORFeome project human cDNA clones) by two bioinformatic N-myristoylation prediction systems, NMT-The MYR Predictor and Myristoylator. After database searches to exclude known human N-myristoylated proteins, 37 cDNA clones were selected as potential human N-myristoylated proteins. The susceptibility of these cDNA clones to protein N-myristoylation was first evaluated using fusion proteins in which the N-terminal ten amino acid residues were fused to an epitope-tagged model protein. Then, protein N-myristoylation of the gene products of full-length cDNAs was evaluated by metabolic labeling experiments both in an insect cell-free protein synthesis system and in transfected human cells. As a result, the products of 13 cDNA clones (FBXL7, PPM1B, SAMM50, PLEKHN, AIFM3, C22orf42, STK32A, FAM131C, DRICH1, MCC1, HID1, P2RX5, STK32B) were found to be human N-myristoylated proteins. Analysis of the role of protein N-myristoylation on the intracellular localization of SAMM50, a mitochondrial outer membrane protein, revealed that protein N-myristoylation was required for proper targeting of SAMM50 to mitochondria. Thus, the strategy used in this study is useful for the identification of physiologically important human N-myristoylated proteins from human cDNA resources.
authors: Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Yamaguchi, 753-8515, Japan.
authors: Emi
authors: E
authors: Takamitsu
authors: Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Yamaguchi, 753-8515, Japan.
authors: Motoaki
authors: M
authors: Otsuka
authors: Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Yamaguchi, 753-8515, Japan.
authors: Tatsuki
authors: T
authors: Haebara
authors: Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Yamaguchi, 753-8515, Japan.
authors: Manami
authors: M
authors: Yano
authors: Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Yamaguchi, 753-8515, Japan.
authors: Kanako
authors: K
authors: Matsuzaki
authors: Department of Cell Chemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, 700-8558, Japan.
authors: Hirotsugu
authors: H
authors: Kobuchi
authors: Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Yamaguchi, 753-8515, Japan.
authors: Koko
authors: K
authors: Moriya
authors: Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Yamaguchi, 753-8515, Japan; Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, 753-8515, Japan.
authors: Toshihiko
authors: T
authors: Utsumi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002474:Cell-Free System; D002522:Chlorocebus aethiops; D018076:DNA, Complementary / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism*; D008969:Molecular Sequence Data; D019814:Myristic Acid / Q000737:chemistry*; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D017386:Sequence Homology, Amino Acid; D014162:Transfection
chemical_list: D018076:DNA, Complementary; D008565:Membrane Proteins; D019814:Myristic Acid