Transmembrane topology and oligomeric nature of an astrocytic membrane protein, MLC1
MLC1 is a membrane protein mainly expressed in astrocytes, and genetic mutations lead to the development of a leukodystrophy, megalencephalic leukoencephalopathy with subcortical cysts disease. Currently, the biochemical properties of the MLC1 protein are largely unknown. In this study, we aimed to...
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| Format: | Article |
| Language: | English |
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The Royal Society
2021-12-01
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| Series: | Open Biology |
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| Online Access: | https://royalsocietypublishing.org/doi/10.1098/rsob.210103 |
| _version_ | 1797837557688434688 |
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| author | Junmo Hwang Kunwoong Park Ga-Young Lee Bo Young Yoon Hyunmin Kim Sung Hoon Roh Byoung-Cheol Lee Kipom Kim Hyun-Ho Lim |
| author_facet | Junmo Hwang Kunwoong Park Ga-Young Lee Bo Young Yoon Hyunmin Kim Sung Hoon Roh Byoung-Cheol Lee Kipom Kim Hyun-Ho Lim |
| author_sort | Junmo Hwang |
| collection | DOAJ |
| description | MLC1 is a membrane protein mainly expressed in astrocytes, and genetic mutations lead to the development of a leukodystrophy, megalencephalic leukoencephalopathy with subcortical cysts disease. Currently, the biochemical properties of the MLC1 protein are largely unknown. In this study, we aimed to characterize the transmembrane (TM) topology and oligomeric nature of the MLC1 protein. Systematic immunofluorescence staining data revealed that the MLC1 protein has eight TM domains and that both the N- and C-terminus face the cytoplasm. We found that MLC1 can be purified as an oligomer and could form a trimeric complex in both detergent micelles and reconstituted proteoliposomes. Additionally, a single-molecule photobleaching experiment showed that MLC1 protein complexes could consist of three MLC1 monomers in the reconstituted proteoliposomes. These results can provide a basis for both the high-resolution structural determination and functional characterization of the MLC1 protein. |
| first_indexed | 2024-04-09T15:27:44Z |
| format | Article |
| id | doaj.art-38f4b72b65bd4330b9ced569bc801818 |
| institution | Directory Open Access Journal |
| issn | 2046-2441 |
| language | English |
| last_indexed | 2024-04-09T15:27:44Z |
| publishDate | 2021-12-01 |
| publisher | The Royal Society |
| record_format | Article |
| series | Open Biology |
| spelling | doaj.art-38f4b72b65bd4330b9ced569bc8018182023-04-28T11:06:27ZengThe Royal SocietyOpen Biology2046-24412021-12-01111210.1098/rsob.210103Transmembrane topology and oligomeric nature of an astrocytic membrane protein, MLC1Junmo Hwang0Kunwoong Park1Ga-Young Lee2Bo Young Yoon3Hyunmin Kim4Sung Hoon Roh5Byoung-Cheol Lee6Kipom Kim7Hyun-Ho Lim8Neurovascular Unit Research Group, Korea Brain Research Institute (KBRI), 61 Cheomdan-ro, Dong-gu, Daegu 41068, Republic of KoreaNeurovascular Unit Research Group, Korea Brain Research Institute (KBRI), 61 Cheomdan-ro, Dong-gu, Daegu 41068, Republic of KoreaBrain Research Core Facility, Korea Brain Research Institute (KBRI), Daegu, Republic of KoreaNeurovascular Unit Research Group, Korea Brain Research Institute (KBRI), 61 Cheomdan-ro, Dong-gu, Daegu 41068, Republic of KoreaSchool of Biological Science, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, Republic of KoreaSchool of Biological Science, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, Republic of KoreaNeurovascular Unit Research Group, Korea Brain Research Institute (KBRI), 61 Cheomdan-ro, Dong-gu, Daegu 41068, Republic of KoreaBrain Research Core Facility, Korea Brain Research Institute (KBRI), Daegu, Republic of KoreaNeurovascular Unit Research Group, Korea Brain Research Institute (KBRI), 61 Cheomdan-ro, Dong-gu, Daegu 41068, Republic of KoreaMLC1 is a membrane protein mainly expressed in astrocytes, and genetic mutations lead to the development of a leukodystrophy, megalencephalic leukoencephalopathy with subcortical cysts disease. Currently, the biochemical properties of the MLC1 protein are largely unknown. In this study, we aimed to characterize the transmembrane (TM) topology and oligomeric nature of the MLC1 protein. Systematic immunofluorescence staining data revealed that the MLC1 protein has eight TM domains and that both the N- and C-terminus face the cytoplasm. We found that MLC1 can be purified as an oligomer and could form a trimeric complex in both detergent micelles and reconstituted proteoliposomes. Additionally, a single-molecule photobleaching experiment showed that MLC1 protein complexes could consist of three MLC1 monomers in the reconstituted proteoliposomes. These results can provide a basis for both the high-resolution structural determination and functional characterization of the MLC1 protein.https://royalsocietypublishing.org/doi/10.1098/rsob.210103MLC1membrane proteintransmembrane topologysubunit stoichiometryfluorescence quenching |
| spellingShingle | Junmo Hwang Kunwoong Park Ga-Young Lee Bo Young Yoon Hyunmin Kim Sung Hoon Roh Byoung-Cheol Lee Kipom Kim Hyun-Ho Lim Transmembrane topology and oligomeric nature of an astrocytic membrane protein, MLC1 Open Biology MLC1 membrane protein transmembrane topology subunit stoichiometry fluorescence quenching |
| title | Transmembrane topology and oligomeric nature of an astrocytic membrane protein, MLC1 |
| title_full | Transmembrane topology and oligomeric nature of an astrocytic membrane protein, MLC1 |
| title_fullStr | Transmembrane topology and oligomeric nature of an astrocytic membrane protein, MLC1 |
| title_full_unstemmed | Transmembrane topology and oligomeric nature of an astrocytic membrane protein, MLC1 |
| title_short | Transmembrane topology and oligomeric nature of an astrocytic membrane protein, MLC1 |
| title_sort | transmembrane topology and oligomeric nature of an astrocytic membrane protein mlc1 |
| topic | MLC1 membrane protein transmembrane topology subunit stoichiometry fluorescence quenching |
| url | https://royalsocietypublishing.org/doi/10.1098/rsob.210103 |
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