Analysis of Dysferlin Direct Interactions with Putative Repair Proteins Links Apoptotic Signaling to Ca<sup>2+</sup> Elevation via PDCD6 and FKBP8
Quantitative surface plasmon resonance (SPR) was utilized to determine binding strength and calcium dependence of direct interactions between dysferlin and proteins likely to mediate skeletal muscle repair, interrupted in limb girdle muscular dystrophy type 2B/R2. Dysferlin canonical C2A (cC2A) and...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/5/4707 |
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| author | Dennis G. Drescher Marian J. Drescher Dakshnamurthy Selvakumar Neeraja P. Annam |
| author_facet | Dennis G. Drescher Marian J. Drescher Dakshnamurthy Selvakumar Neeraja P. Annam |
| author_sort | Dennis G. Drescher |
| collection | DOAJ |
| description | Quantitative surface plasmon resonance (SPR) was utilized to determine binding strength and calcium dependence of direct interactions between dysferlin and proteins likely to mediate skeletal muscle repair, interrupted in limb girdle muscular dystrophy type 2B/R2. Dysferlin canonical C2A (cC2A) and C2F/G domains directly interacted with annexin A1, calpain-3, caveolin-3, affixin, AHNAK1, syntaxin-4, and mitsugumin-53, with cC2A the primary target and C2F lesser involved, overall demonstrating positive calcium dependence. Dysferlin C2 pairings alone showed negative calcium dependence in almost all cases. Like otoferlin, dysferlin directly interacted via its carboxy terminus with FKBP8, an anti-apoptotic outer mitochondrial membrane protein, and via its C2DE domain with apoptosis-linked gene (ALG-2/PDCD6), linking anti-apoptosis with apoptosis. Confocal Z-stack immunofluorescence confirmed co-compartmentalization of PDCD6 and FKBP8 at the sarcolemmal membrane. Our evidence supports the hypothesis that prior to injury, dysferlin C2 domains self-interact and give rise to a folded, compact structure as indicated for otoferlin. With elevation of intracellular Ca<sup>2+</sup> in injury, dysferlin would unfold and expose the cC2A domain for interaction with annexin A1, calpain-3, mitsugumin 53, affixin, and caveolin-3, and dysferlin would realign from its interactions with PDCD6 at basal calcium levels to interact strongly with FKBP8, an intramolecular rearrangement facilitating membrane repair. |
| first_indexed | 2024-03-11T07:23:06Z |
| format | Article |
| id | doaj.art-ed26fc41cbd04181a0979981fcfaa457 |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T07:23:06Z |
| publishDate | 2023-02-01 |
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| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-ed26fc41cbd04181a0979981fcfaa4572023-11-17T07:52:16ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-02-01245470710.3390/ijms24054707Analysis of Dysferlin Direct Interactions with Putative Repair Proteins Links Apoptotic Signaling to Ca<sup>2+</sup> Elevation via PDCD6 and FKBP8Dennis G. Drescher0Marian J. Drescher1Dakshnamurthy Selvakumar2Neeraja P. Annam3Laboratory of Bio-otology, Department of Otolaryngology, Wayne State University School of Medicine, Detroit, MI 48201, USALaboratory of Bio-otology, Department of Otolaryngology, Wayne State University School of Medicine, Detroit, MI 48201, USALaboratory of Bio-otology, Department of Otolaryngology, Wayne State University School of Medicine, Detroit, MI 48201, USALaboratory of Bio-otology, Department of Otolaryngology, Wayne State University School of Medicine, Detroit, MI 48201, USAQuantitative surface plasmon resonance (SPR) was utilized to determine binding strength and calcium dependence of direct interactions between dysferlin and proteins likely to mediate skeletal muscle repair, interrupted in limb girdle muscular dystrophy type 2B/R2. Dysferlin canonical C2A (cC2A) and C2F/G domains directly interacted with annexin A1, calpain-3, caveolin-3, affixin, AHNAK1, syntaxin-4, and mitsugumin-53, with cC2A the primary target and C2F lesser involved, overall demonstrating positive calcium dependence. Dysferlin C2 pairings alone showed negative calcium dependence in almost all cases. Like otoferlin, dysferlin directly interacted via its carboxy terminus with FKBP8, an anti-apoptotic outer mitochondrial membrane protein, and via its C2DE domain with apoptosis-linked gene (ALG-2/PDCD6), linking anti-apoptosis with apoptosis. Confocal Z-stack immunofluorescence confirmed co-compartmentalization of PDCD6 and FKBP8 at the sarcolemmal membrane. Our evidence supports the hypothesis that prior to injury, dysferlin C2 domains self-interact and give rise to a folded, compact structure as indicated for otoferlin. With elevation of intracellular Ca<sup>2+</sup> in injury, dysferlin would unfold and expose the cC2A domain for interaction with annexin A1, calpain-3, mitsugumin 53, affixin, and caveolin-3, and dysferlin would realign from its interactions with PDCD6 at basal calcium levels to interact strongly with FKBP8, an intramolecular rearrangement facilitating membrane repair.https://www.mdpi.com/1422-0067/24/5/4707calcium dependenceC2 domainsdysferlinFKBP8limb girdle muscular dystrophy type 2B/R2PDCD6 |
| spellingShingle | Dennis G. Drescher Marian J. Drescher Dakshnamurthy Selvakumar Neeraja P. Annam Analysis of Dysferlin Direct Interactions with Putative Repair Proteins Links Apoptotic Signaling to Ca<sup>2+</sup> Elevation via PDCD6 and FKBP8 International Journal of Molecular Sciences calcium dependence C2 domains dysferlin FKBP8 limb girdle muscular dystrophy type 2B/R2 PDCD6 |
| title | Analysis of Dysferlin Direct Interactions with Putative Repair Proteins Links Apoptotic Signaling to Ca<sup>2+</sup> Elevation via PDCD6 and FKBP8 |
| title_full | Analysis of Dysferlin Direct Interactions with Putative Repair Proteins Links Apoptotic Signaling to Ca<sup>2+</sup> Elevation via PDCD6 and FKBP8 |
| title_fullStr | Analysis of Dysferlin Direct Interactions with Putative Repair Proteins Links Apoptotic Signaling to Ca<sup>2+</sup> Elevation via PDCD6 and FKBP8 |
| title_full_unstemmed | Analysis of Dysferlin Direct Interactions with Putative Repair Proteins Links Apoptotic Signaling to Ca<sup>2+</sup> Elevation via PDCD6 and FKBP8 |
| title_short | Analysis of Dysferlin Direct Interactions with Putative Repair Proteins Links Apoptotic Signaling to Ca<sup>2+</sup> Elevation via PDCD6 and FKBP8 |
| title_sort | analysis of dysferlin direct interactions with putative repair proteins links apoptotic signaling to ca sup 2 sup elevation via pdcd6 and fkbp8 |
| topic | calcium dependence C2 domains dysferlin FKBP8 limb girdle muscular dystrophy type 2B/R2 PDCD6 |
| url | https://www.mdpi.com/1422-0067/24/5/4707 |
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