Identification of Human N-Myristoylated Proteins from Human Complementary DNA Resources by Cell-Free and Cellular Metabolic Labeling Analyses
| Autor: | Kanako Matsuzaki, Emi Takamitsu, Hirotsugu Kobuchi, Tatsuki Haebara, Toshihiko Utsumi, Manami Yano, Koko Moriya, Motoaki Otsuka |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
DNA
Complementary Molecular Sequence Data lcsh:Medicine Biology Transfection Myristic Acid Complementary DNA Chlorocebus aethiops Protein biosynthesis Animals Humans Amino Acid Sequence ORFeome lcsh:Science Peptide sequence Integral membrane protein Cell-free protein synthesis Multidisciplinary Cell-Free System Sequence Homology Amino Acid lcsh:R Membrane Proteins Fusion protein Molecular biology Mitochondria HEK293 Cells Membrane protein Biochemistry Microscopy Fluorescence Protein Biosynthesis COS Cells lcsh:Q lipids (amino acids peptides and proteins) Protein Processing Post-Translational Research Article |
| Zdroj: | PLoS ONE PLoS ONE, Vol 10, Iss 8, p e0136360 (2015) |
| ISSN: | 1932-6203 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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