GnT1IP-L specifically inhibits MGAT1 in the Golgi via its luminal domain
Mouse GnT1IP-L, and membrane-bound GnT1IP-S (MGAT4D) expressed in cultured cells inhibit MGAT1, the N-acetylglucosaminyltransferase that initiates the synthesis of hybrid and complex N-glycans. However, it is not known where in the secretory pathway GnT1IP-L inhibits MGAT1, nor whether GnT1IP-L inhi...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2015-09-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/08916 |
| _version_ | 1811235837773348864 |
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| author | Hung-Hsiang Huang Antti Hassinen Subha Sundaram Andrej-Nikolai Spiess Sakari Kellokumpu Pamela Stanley |
| author_facet | Hung-Hsiang Huang Antti Hassinen Subha Sundaram Andrej-Nikolai Spiess Sakari Kellokumpu Pamela Stanley |
| author_sort | Hung-Hsiang Huang |
| collection | DOAJ |
| description | Mouse GnT1IP-L, and membrane-bound GnT1IP-S (MGAT4D) expressed in cultured cells inhibit MGAT1, the N-acetylglucosaminyltransferase that initiates the synthesis of hybrid and complex N-glycans. However, it is not known where in the secretory pathway GnT1IP-L inhibits MGAT1, nor whether GnT1IP-L inhibits other N-glycan branching N-acetylglucosaminyltransferases of the medial Golgi. We show here that the luminal domain of GnT1IP-L contains its inhibitory activity. Retention of GnT1IP-L in the endoplasmic reticulum (ER) via the N-terminal region of human invariant chain p33, with or without C-terminal KDEL, markedly reduced inhibitory activity. Dynamic fluorescent resonance energy transfer (FRET) and bimolecular fluorescence complementation (BiFC) assays revealed homomeric interactions for GnT1IP-L in the ER, and heteromeric interactions with MGAT1 in the Golgi. GnT1IP-L did not generate a FRET signal with MGAT2, MGAT3, MGAT4B or MGAT5 medial Golgi GlcNAc-tranferases. GnT1IP/Mgat4d transcripts are expressed predominantly in spermatocytes and spermatids in mouse, and are reduced in men with impaired spermatogenesis. |
| first_indexed | 2024-04-12T11:58:50Z |
| format | Article |
| id | doaj.art-3b94237a855143d78bc6f1f35620657b |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T11:58:50Z |
| publishDate | 2015-09-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-3b94237a855143d78bc6f1f35620657b2022-12-22T03:33:54ZengeLife Sciences Publications LtdeLife2050-084X2015-09-01410.7554/eLife.08916GnT1IP-L specifically inhibits MGAT1 in the Golgi via its luminal domainHung-Hsiang Huang0Antti Hassinen1Subha Sundaram2Andrej-Nikolai Spiess3Sakari Kellokumpu4Pamela Stanley5Department of Cell Biology, Albert Einstein College of Medicine, New York, United StatesFaculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, FinlandDepartment of Cell Biology, Albert Einstein College of Medicine, New York, United StatesAndrology Division, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyFaculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, FinlandDepartment of Cell Biology, Albert Einstein College of Medicine, New York, United StatesMouse GnT1IP-L, and membrane-bound GnT1IP-S (MGAT4D) expressed in cultured cells inhibit MGAT1, the N-acetylglucosaminyltransferase that initiates the synthesis of hybrid and complex N-glycans. However, it is not known where in the secretory pathway GnT1IP-L inhibits MGAT1, nor whether GnT1IP-L inhibits other N-glycan branching N-acetylglucosaminyltransferases of the medial Golgi. We show here that the luminal domain of GnT1IP-L contains its inhibitory activity. Retention of GnT1IP-L in the endoplasmic reticulum (ER) via the N-terminal region of human invariant chain p33, with or without C-terminal KDEL, markedly reduced inhibitory activity. Dynamic fluorescent resonance energy transfer (FRET) and bimolecular fluorescence complementation (BiFC) assays revealed homomeric interactions for GnT1IP-L in the ER, and heteromeric interactions with MGAT1 in the Golgi. GnT1IP-L did not generate a FRET signal with MGAT2, MGAT3, MGAT4B or MGAT5 medial Golgi GlcNAc-tranferases. GnT1IP/Mgat4d transcripts are expressed predominantly in spermatocytes and spermatids in mouse, and are reduced in men with impaired spermatogenesis.https://elifesciences.org/articles/08916GnT1IP-LMGAT1 inhibitorcomplex N-glycansGolgi interactionspermatogenesis |
| spellingShingle | Hung-Hsiang Huang Antti Hassinen Subha Sundaram Andrej-Nikolai Spiess Sakari Kellokumpu Pamela Stanley GnT1IP-L specifically inhibits MGAT1 in the Golgi via its luminal domain eLife GnT1IP-L MGAT1 inhibitor complex N-glycans Golgi interaction spermatogenesis |
| title | GnT1IP-L specifically inhibits MGAT1 in the Golgi via its luminal domain |
| title_full | GnT1IP-L specifically inhibits MGAT1 in the Golgi via its luminal domain |
| title_fullStr | GnT1IP-L specifically inhibits MGAT1 in the Golgi via its luminal domain |
| title_full_unstemmed | GnT1IP-L specifically inhibits MGAT1 in the Golgi via its luminal domain |
| title_short | GnT1IP-L specifically inhibits MGAT1 in the Golgi via its luminal domain |
| title_sort | gnt1ip l specifically inhibits mgat1 in the golgi via its luminal domain |
| topic | GnT1IP-L MGAT1 inhibitor complex N-glycans Golgi interaction spermatogenesis |
| url | https://elifesciences.org/articles/08916 |
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