ANKRD22 is an N-myristoylated hairpin-like monotopic membrane protein specifically localized to lipid droplets
Abstract The membrane topology and intracellular localization of ANKRD22, a novel human N-myristoylated protein with a predicted single-pass transmembrane domain that was recently reported to be overexpressed in cancer, were examined. Immunofluorescence staining of COS-1 cells transfected with cDNA...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-09-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-98486-8 |
| _version_ | 1818342347597938688 |
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| author | Toshihiko Utsumi Takuro Hosokawa Mayu Shichita Misato Nishiue Natsuko Iwamoto Haruna Harada Aya Kiwado Manami Yano Motoaki Otsuka Koko Moriya |
| author_facet | Toshihiko Utsumi Takuro Hosokawa Mayu Shichita Misato Nishiue Natsuko Iwamoto Haruna Harada Aya Kiwado Manami Yano Motoaki Otsuka Koko Moriya |
| author_sort | Toshihiko Utsumi |
| collection | DOAJ |
| description | Abstract The membrane topology and intracellular localization of ANKRD22, a novel human N-myristoylated protein with a predicted single-pass transmembrane domain that was recently reported to be overexpressed in cancer, were examined. Immunofluorescence staining of COS-1 cells transfected with cDNA encoding ANKRD22 coupled with organelle markers revealed that ANKRD22 localized specifically to lipid droplets (LD). Analysis of the intracellular localization of ANKRD22 mutants C-terminally fused to glycosylatable tumor necrosis factor (GLCTNF) and assessment of their susceptibility to protein N-glycosylation revealed that ANKRD22 is synthesized on the endoplasmic reticulum (ER) membrane as an N-myristoylated hairpin-like monotopic membrane protein with the amino- and carboxyl termini facing the cytoplasm and then sorted to LD. Pro98 located at the center of the predicted membrane domain was found to be essential for the formation of the hairpin-like monotopic topology of ANKRD22. Moreover, the hairpin-like monotopic topology, and positively charged residues located near the C-terminus were demonstrated to be required for the sorting of ANKRD22 from ER to LD. Protein N-myristoylation was found to positively affect the LD localization. Thus, multiple factors, including hairpin-like monotopic membrane topology, C-terminal positively charged residues, and protein N-myristoylation cooperatively affected the intracellular targeting of ANKRD22 to LD. |
| first_indexed | 2024-12-13T16:13:15Z |
| format | Article |
| id | doaj.art-c686d4478e2547978727bd554bb80151 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-13T16:13:15Z |
| publishDate | 2021-09-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-c686d4478e2547978727bd554bb801512022-12-21T23:38:53ZengNature PortfolioScientific Reports2045-23222021-09-0111111710.1038/s41598-021-98486-8ANKRD22 is an N-myristoylated hairpin-like monotopic membrane protein specifically localized to lipid dropletsToshihiko Utsumi0Takuro Hosokawa1Mayu Shichita2Misato Nishiue3Natsuko Iwamoto4Haruna Harada5Aya Kiwado6Manami Yano7Motoaki Otsuka8Koko Moriya9Graduate School of Sciences and Technology for Innovation, Yamaguchi UniversityGraduate School of Sciences and Technology for Innovation, Yamaguchi UniversityGraduate School of Sciences and Technology for Innovation, Yamaguchi UniversityGraduate School of Sciences and Technology for Innovation, Yamaguchi UniversityGraduate School of Sciences and Technology for Innovation, Yamaguchi UniversityGraduate School of Sciences and Technology for Innovation, Yamaguchi UniversityGraduate School of Sciences and Technology for Innovation, Yamaguchi UniversityGraduate School of Sciences and Technology for Innovation, Yamaguchi UniversityGraduate School of Sciences and Technology for Innovation, Yamaguchi UniversityGraduate School of Sciences and Technology for Innovation, Yamaguchi UniversityAbstract The membrane topology and intracellular localization of ANKRD22, a novel human N-myristoylated protein with a predicted single-pass transmembrane domain that was recently reported to be overexpressed in cancer, were examined. Immunofluorescence staining of COS-1 cells transfected with cDNA encoding ANKRD22 coupled with organelle markers revealed that ANKRD22 localized specifically to lipid droplets (LD). Analysis of the intracellular localization of ANKRD22 mutants C-terminally fused to glycosylatable tumor necrosis factor (GLCTNF) and assessment of their susceptibility to protein N-glycosylation revealed that ANKRD22 is synthesized on the endoplasmic reticulum (ER) membrane as an N-myristoylated hairpin-like monotopic membrane protein with the amino- and carboxyl termini facing the cytoplasm and then sorted to LD. Pro98 located at the center of the predicted membrane domain was found to be essential for the formation of the hairpin-like monotopic topology of ANKRD22. Moreover, the hairpin-like monotopic topology, and positively charged residues located near the C-terminus were demonstrated to be required for the sorting of ANKRD22 from ER to LD. Protein N-myristoylation was found to positively affect the LD localization. Thus, multiple factors, including hairpin-like monotopic membrane topology, C-terminal positively charged residues, and protein N-myristoylation cooperatively affected the intracellular targeting of ANKRD22 to LD.https://doi.org/10.1038/s41598-021-98486-8 |
| spellingShingle | Toshihiko Utsumi Takuro Hosokawa Mayu Shichita Misato Nishiue Natsuko Iwamoto Haruna Harada Aya Kiwado Manami Yano Motoaki Otsuka Koko Moriya ANKRD22 is an N-myristoylated hairpin-like monotopic membrane protein specifically localized to lipid droplets Scientific Reports |
| title | ANKRD22 is an N-myristoylated hairpin-like monotopic membrane protein specifically localized to lipid droplets |
| title_full | ANKRD22 is an N-myristoylated hairpin-like monotopic membrane protein specifically localized to lipid droplets |
| title_fullStr | ANKRD22 is an N-myristoylated hairpin-like monotopic membrane protein specifically localized to lipid droplets |
| title_full_unstemmed | ANKRD22 is an N-myristoylated hairpin-like monotopic membrane protein specifically localized to lipid droplets |
| title_short | ANKRD22 is an N-myristoylated hairpin-like monotopic membrane protein specifically localized to lipid droplets |
| title_sort | ankrd22 is an n myristoylated hairpin like monotopic membrane protein specifically localized to lipid droplets |
| url | https://doi.org/10.1038/s41598-021-98486-8 |
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