A systematic immunoprecipitation approach reinforces the concept of common conformational alterations in amyotrophic lateral sclerosis-linked SOD1 mutants
Mutations in the Cu, Zn superoxide dismutase (SOD1) gene are one of the causative agents of amyotrophic lateral sclerosis (ALS). Although more than 100 different mutations in SOD1 have been identified, it is unclear whether all the mutations are pathogenic or just single nucleotide polymorphisms (SN...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015-10-01
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| Series: | Neurobiology of Disease |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0969996115300310 |
| _version_ | 1818651604366131200 |
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| author | Takao Fujisawa Namiko Yamaguchi Hisae Kadowaki Yuka Tsukamoto Naomi Tsuburaya Atsushi Tsubota Hiromitsu Takahashi Isao Naguro Yuji Takahashi Jun Goto Shoji Tsuji Hideki Nishitoh Kengo Homma Hidenori Ichijo |
| author_facet | Takao Fujisawa Namiko Yamaguchi Hisae Kadowaki Yuka Tsukamoto Naomi Tsuburaya Atsushi Tsubota Hiromitsu Takahashi Isao Naguro Yuji Takahashi Jun Goto Shoji Tsuji Hideki Nishitoh Kengo Homma Hidenori Ichijo |
| author_sort | Takao Fujisawa |
| collection | DOAJ |
| description | Mutations in the Cu, Zn superoxide dismutase (SOD1) gene are one of the causative agents of amyotrophic lateral sclerosis (ALS). Although more than 100 different mutations in SOD1 have been identified, it is unclear whether all the mutations are pathogenic or just single nucleotide polymorphisms (SNPs) unrelated to the disease. Our previous systematic analysis found that all pathogenic SOD1 mutants (SOD1mut) have a common property, namely, an association with Derlin-1, a component of the endoplasmic reticulum-associated degradation machinery. For the proposed mechanism, we found that most pathogenic SOD1mut have a constitutively exposed Derlin-1-binding region (DBR), which is concealed in wild-type SOD1 (SOD1WT). Moreover, we generated MS785, a monoclonal antibody against DBR. MS785 distinguished most ALS-causative SOD1mut from both SOD1WT and non-toxic SOD1mut. However, MS785 could not recognize SOD1mut that has mutations in the MS785 epitope region. Here, we developed a new diagnostic antibody, which could compensate for this shortcoming of MS785. We hypothesized that in ALS-causative SOD1mut, the DBR-neighboring region [SOD1(30–40)] may also be exposed. We then generated MS27, a monoclonal antibody against SOD1(30–40). We found that MS27 could distinguish SOD1WT from the pathogenic SOD1mut, which has mutations in the MS785 epitope region. Moreover, all pathogenic SOD1mut, without exception, were immunoprecipitated with a combination of MS785 and MS27. The MS785–MS27 combination could be developed as a novel mechanism-based biomarker for the diagnosis of ALS. |
| first_indexed | 2024-12-17T02:08:45Z |
| format | Article |
| id | doaj.art-75c61bf9da0c485e922c8c413b437906 |
| institution | Directory Open Access Journal |
| issn | 1095-953X |
| language | English |
| last_indexed | 2024-12-17T02:08:45Z |
| publishDate | 2015-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neurobiology of Disease |
| spelling | doaj.art-75c61bf9da0c485e922c8c413b4379062022-12-21T22:07:38ZengElsevierNeurobiology of Disease1095-953X2015-10-0182478486A systematic immunoprecipitation approach reinforces the concept of common conformational alterations in amyotrophic lateral sclerosis-linked SOD1 mutantsTakao Fujisawa0Namiko Yamaguchi1Hisae Kadowaki2Yuka Tsukamoto3Naomi Tsuburaya4Atsushi Tsubota5Hiromitsu Takahashi6Isao Naguro7Yuji Takahashi8Jun Goto9Shoji Tsuji10Hideki Nishitoh11Kengo Homma12Hidenori Ichijo13Laboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanLaboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanSection of Biochemistry and Molecular Biology, Department of Medical Sciences, Miyazaki Medical College, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki 889-1692, JapanLaboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanLaboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanLaboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanLaboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanLaboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanDepartment of Neurology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, JapanDepartment of Neurology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, JapanDepartment of Neurology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, JapanSection of Biochemistry and Molecular Biology, Department of Medical Sciences, Miyazaki Medical College, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki 889-1692, JapanLaboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanLaboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan; Corresponding author at: Laboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.Mutations in the Cu, Zn superoxide dismutase (SOD1) gene are one of the causative agents of amyotrophic lateral sclerosis (ALS). Although more than 100 different mutations in SOD1 have been identified, it is unclear whether all the mutations are pathogenic or just single nucleotide polymorphisms (SNPs) unrelated to the disease. Our previous systematic analysis found that all pathogenic SOD1 mutants (SOD1mut) have a common property, namely, an association with Derlin-1, a component of the endoplasmic reticulum-associated degradation machinery. For the proposed mechanism, we found that most pathogenic SOD1mut have a constitutively exposed Derlin-1-binding region (DBR), which is concealed in wild-type SOD1 (SOD1WT). Moreover, we generated MS785, a monoclonal antibody against DBR. MS785 distinguished most ALS-causative SOD1mut from both SOD1WT and non-toxic SOD1mut. However, MS785 could not recognize SOD1mut that has mutations in the MS785 epitope region. Here, we developed a new diagnostic antibody, which could compensate for this shortcoming of MS785. We hypothesized that in ALS-causative SOD1mut, the DBR-neighboring region [SOD1(30–40)] may also be exposed. We then generated MS27, a monoclonal antibody against SOD1(30–40). We found that MS27 could distinguish SOD1WT from the pathogenic SOD1mut, which has mutations in the MS785 epitope region. Moreover, all pathogenic SOD1mut, without exception, were immunoprecipitated with a combination of MS785 and MS27. The MS785–MS27 combination could be developed as a novel mechanism-based biomarker for the diagnosis of ALS.http://www.sciencedirect.com/science/article/pii/S0969996115300310Amyotrophic lateral sclerosisMutant SOD1ER stressMutant SOD1-specific antibodyELISACommon conformational change |
| spellingShingle | Takao Fujisawa Namiko Yamaguchi Hisae Kadowaki Yuka Tsukamoto Naomi Tsuburaya Atsushi Tsubota Hiromitsu Takahashi Isao Naguro Yuji Takahashi Jun Goto Shoji Tsuji Hideki Nishitoh Kengo Homma Hidenori Ichijo A systematic immunoprecipitation approach reinforces the concept of common conformational alterations in amyotrophic lateral sclerosis-linked SOD1 mutants Neurobiology of Disease Amyotrophic lateral sclerosis Mutant SOD1 ER stress Mutant SOD1-specific antibody ELISA Common conformational change |
| title | A systematic immunoprecipitation approach reinforces the concept of common conformational alterations in amyotrophic lateral sclerosis-linked SOD1 mutants |
| title_full | A systematic immunoprecipitation approach reinforces the concept of common conformational alterations in amyotrophic lateral sclerosis-linked SOD1 mutants |
| title_fullStr | A systematic immunoprecipitation approach reinforces the concept of common conformational alterations in amyotrophic lateral sclerosis-linked SOD1 mutants |
| title_full_unstemmed | A systematic immunoprecipitation approach reinforces the concept of common conformational alterations in amyotrophic lateral sclerosis-linked SOD1 mutants |
| title_short | A systematic immunoprecipitation approach reinforces the concept of common conformational alterations in amyotrophic lateral sclerosis-linked SOD1 mutants |
| title_sort | systematic immunoprecipitation approach reinforces the concept of common conformational alterations in amyotrophic lateral sclerosis linked sod1 mutants |
| topic | Amyotrophic lateral sclerosis Mutant SOD1 ER stress Mutant SOD1-specific antibody ELISA Common conformational change |
| url | http://www.sciencedirect.com/science/article/pii/S0969996115300310 |
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