Identification of human N-myristoylated proteins from human complementary DNA resources by cell-free and cellular metabolic labeling analyses

Emi Takamitsu, Motoaki Otsuka, Tatsuki Haebara, Manami Yano, Kanako Matsuzaki, Hirotsugu Kobuchi, Koko Moriya, Toshihiko Utsumi

Research output: Contribution to journalArticle

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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 Nterminal 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.

Original languageEnglish
Article numbere0136360
JournalPLoS One
Volume10
Issue number8
DOIs
Publication statusPublished - Aug 26 2015

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Forensic Anthropology
complementary DNA
Labeling
Complementary DNA
Proteins
proteins
cells
Clone Cells
clones
Personnel
outer membrane proteins
Mitochondria
bioinformatics
Bioinformatics
Computational Biology
epitopes
Insects
mitochondria
Epitopes
Membrane Proteins

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

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Identification of human N-myristoylated proteins from human complementary DNA resources by cell-free and cellular metabolic labeling analyses. / Takamitsu, Emi; Otsuka, Motoaki; Haebara, Tatsuki; Yano, Manami; Matsuzaki, Kanako; Kobuchi, Hirotsugu; Moriya, Koko; Utsumi, Toshihiko.

In: PLoS One, Vol. 10, No. 8, e0136360, 26.08.2015.

Research output: Contribution to journalArticle

Takamitsu, Emi ; Otsuka, Motoaki ; Haebara, Tatsuki ; Yano, Manami ; Matsuzaki, Kanako ; Kobuchi, Hirotsugu ; Moriya, Koko ; Utsumi, Toshihiko. / Identification of human N-myristoylated proteins from human complementary DNA resources by cell-free and cellular metabolic labeling analyses. In: PLoS One. 2015 ; Vol. 10, No. 8.
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