Dysferlin Facts

The following Dysferlin antibodies have been published:

• SmaI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• SalI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• BamHI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• HM38: Ho M, et al. 2004. Human Molecular Genetics 13(18):1999-2010
• F4: Huang Y, et al. 2005. European J of Human Genetics 13:721-730
• H7: Huang Y, et al. 2005. European J of Human Genetics 13:721-730
• Dysf1: Davis DB et al., 2000. Human Molecular Genetics 9:217-226

The following antibodies are directed towards the N-terminus of Dysferlin:

• NCL-Hamlet-2: Novocastra Laboratories Ltd.
• SmaI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• BamHI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• F4: Huang Y, et al. 2005. European J of Human Genetics 13:721-730

The following antibodies react with human Dysferlin:

• NCL-Hamlet: Novocastra Laboratories Ltd.
• NCL-Hamlet-2: Novocastra Laboratories Ltd.
• ab15108: Abcam
• C-19: Santa Cruz Biotechnology, Inc.
• E-20: Santa Cruz Biotechnology, Inc.
• SmaI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• SalI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• BamHI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• F4: Huang Y, et al. 2005. European J of Human Genetics 13:721-730
• H7: Huang Y, et al. 2005. European J of Human Genetics 13:721-730

The NCL-Hamlet antibody from Novocastra Laboratories Ltd. reacts with rat, rabbit, hamster, pig and dog Dysferlin besides the mouse and human protein.

The following Dysferlin antibodies are polyclonal:

• ab15108: Abcam
• C-19: Santa Cruz Biotechnology, Inc.
• E-20: Santa Cruz Biotechnology, Inc.
• SmaI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• SalI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• BamHI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• HM38: Ho M, et al. 2004. Human Molecular Genetics 13(18):1999-2010
• DysF1: Davis DB et al., 2000. Human Molecular Genetics 9:217-226

The following antibodies have been successfully used for Dysferlin immunoblotting:

• NCL-Hamlet: Novocastra Laboratories Ltd.
• NCL-Hamlet-2: Novocastra Laboratories Ltd.
• ab15108: Abcam
• C-19: Santa Cruz Biotechnology, Inc.
• E-20: Santa Cruz Biotechnology, Inc.
• SalI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• HM38: Ho M, et al. 2004. Human Molecular Genetics 13(18):1999-2010
• F4: Huang Y, et al. 2005. European J of Human Genetics 13:721-730
• DysF1: Davis DB et al., 2000. Human Molecular Genetics 9:217-226

The following antibodies have been successfully used for Dysferlin IF or IHC:

• NCL-Hamlet: Novocastra Laboratories Ltd.
• NCL-Hamlet-2: Novocastra Laboratories Ltd.
• ab15108: Abcam
• C-19: Santa Cruz Biotechnology, Inc.
• SalI: Matsuda C, et al. 1999. Neurology 53:1119-1122
• F4: Huang Y, et al. 2005. European J of Human Genetics 13:721-730
• H7: Huang Y, et al. 2005. European J of Human Genetics 13:721-730
• DysF1: Davis DB et al., 2000. Human Molecular Genetics 9:217-226

The epitopes for the various Myoferlin antibodies map as follows:

• MYOF3: amino acids 1088-1107 (Doherty KR et al., 2005. Development 132: 5565-5575)
• Myof1: amino acids 1692-1709 (human Myoferlin) (Davis DB et al., 2000. Human Molecular Genetics 9:217-226)
• Myof2: amino acids 928-945 (human Myoferlin) (Davis DB et al., 2000. Human Molecular Genetics 9:217-226)
• Monoclonal Myoferlin Ab: amino acids 326-344 (human Myoferlin) (Jaiswal JK et al., 2007. Traffic 8:77-88)
• Polyclonal Myoferlin Ab: amino acids 655-755 (human Myoferlin) (Abnova Corporation, catalog number: H00026509-A01 )

Based on the sequence of their synthetic peptide, the following antibodies are not predicted to react with Dysferlin by the authors:

• Myof1: Davis DB et al., 2000. Human Molecular Genetics 9:217-226
• Myof2: Davis DB et al., 2000. Human Molecular Genetics 9:217-226

The Myoferlin antibody published in Jaiswal JK et al., 2007. Traffic 8:77-88 is monoclonal.

Full-length mouse dysferlin cDNA construct has been described in: Vafiadaki et al., Neuroreport 12(3): 625-629.

Yes, 3’ HA-tagged human myoferlin has been described in: Bernatchez et al., JBC 282(42):30745-30753

The highest expression of Dysferlin protein is found in skeletal and cardiac muscle, and kidneys. Dysferlin expression has also been reported in satellite cells (SCs) and certain immune cells like monocytes and macrophages.  In addition, a shorter transcript of 4kb is detectable in the brain tissue and faint, lower molecular weight bands can be detected in the brain stem, spinal cord and sciatic nerve (Glover and Brown, Traffic 8: 785-794; deLuna et al., J of Neuropath Exp Neuro 63 (10): 1104-1113; Anderson et al., Human Molecular Genetics 8 (5): 855-861). Recently Dysferlin expression was also detected in human placental microvillus syncytiotrophoblasts (Vandre et al., Biol Reprod 77: 533-542). Dysferlin positive staining is also observed in case of stomach, lung, liver and spleen tissue from rats (Anderson et al., Human Molecular Genetics 8 (5): 855-861).

The coding sequence of Dysferlin is comprised of 55 exons. Two splice variants of Dysferlin have been described, “original” and “variant-1” that are 6907bp and 6543bp in length respectively. Please see below for more information on Dysferlin splice variants.

No human disorders have been attributed to Annexin A1 or A2 deficiency as of yet.

Affixin was shown to interact with dysferlin in mouse and human skeletal muscles by coimmunoprecipitation. The C-terminal intracellular region of Dysferlin was identified as the Affixin binding domain and the CH1 domain of Affixin was identified as Dysferlin-interacting region. 
Matsuda et al., J Neuropathol Exp Neurol 64 (4): 334- 340

No human disorders have been attributed to Affixin deficiency as of yet.

Carboxy-terminal domains of both AHNAK proteins (AHNAK1 and 2) interact directly with C2A-Dysferlin. The N-terminal portion of Myoferlin also interacts with the C-terminus of both AHNAK proteins. 
Huang et al., FASEB J 21 (3): 732-742

No human disorders have been attributed to AHNAK deficiency as of yet.

Dysferlin and Calpain-3 were shown to interact by coimmunoprecipitation in human muscle homogenates. Full-length as well as degradation products of Calpain-3 were shown to interact.
Huang et al., European Journal of Human Genetics 13 (6): 721-730

Primary mutations in Calpain-3 cause LGMD2A.

Caveolin-3 and Dysferlin were shown to interact weakly in normal biopsied muscle by coimmunoprecipitation. Seven sites in Dysferlin correspond to micro-domains believed to bind the caveolin-3 scaffolding region. 
Matsuda et al., Human Molecular Genetics 10 (17): 1761-1766

Caveolins are required for Dysferlin trafficking of Caveolin-1 or Caveolin-3 mutants which cause accumulation of Dysferlin in the Golgi complex. 
Hernandez-Deviez et al., Human Molecular Genetics 15 (1): 129-142

Caveolin-3 and Dysferlin show only limited co-localization at the sarcolemma in mature muscle fibers and Dysferlin is not particularly enriched in caveolae. It was suggested that the weak association between the two proteins may occur during dysferlin trafficking but not at the membrane. 
Hernandez-Deviez et al., Human Molecular Genetics 15 (1): 129-142

Mutations in Caveolin-3 are associated with numerous muscle pathologies - autosomal dominant LGMD1C, distal myopathy, hyperCKemia and rippling muscle disease (RMD).

A structure for the Dysferlin protein has not yet been elucidated. However, several domains have been identified based on sequence homology to other proteins. Please click here to see a color-coded domain-distribution of Dysferlin.

The SJL/J mice are homozygous for a missense mutation in dysferlin. The mutation is in the 3’ splice junction of Exon 45, which causes a deletion of 171 base pairs corresponding to amino acids 1628-1685 of the Dysferlin protein. This deletion removes a part of the fifth C2 domain (C2E) of the protein. SJL mice have partial Dysferlin expression; expression has been reported to be about 15% of the WT level. In addition, Dysferlin is mislocalized, and is spread throughout the muscle cells rather than being concentrated at the sarcolemma. Dysferlin expression by the C57BL10.SJL-Dysf strain is likely to be similar to that seen in SJL mice.

The A/J mouse has an ETn retrotransposon (5-6kb) inserted in Intron 4 of the dysferlin gene. The mice are homozygous for this insertion. The mice do not express any dysferlin. Similarly, dysferlin expression in the B6.A-Dysf^PRMDGene/J and C57BL/6J-Chr6^A/J/NaJ mice strains is likely to be absent.

In the 129.Dysf^tm1Kcam/J and  B6.129-Dysf^tm1Kcam/J mice, a 12-kb region of the genome, containing the last three exons (Exons 53-55, aa1983-2080) of dysferlin, is deleted. This deletion removes the transmembrane domain of the protein. 129.Dysf^tm1Kcam/J mice are of a mixed 129SvJ and C57BL/6 background and B6.129-Dysf^tm1Kcam/J  mice are on a C57BL/6 background. Both strains are homozygous for this deletion and do not express any dysferlin.

Since the mutations found in A/J and SJL/J mice are naturally occurring, choosing an appropriate control strain is challenging. The SWR/J strain has been used as a dysferlin sufficient control for the SJL/J strain and the A/WySnJ or A/HeJ strains used as dysferlin sufficient controls for the A/J strain. It is important to recognize that the SWR, A/HeJ and A/WySnJ strains are only genetically similar to, and NOT true congenic controls for, either SJL or A/J animals. It is also important to note that at some foreign repositories in Europe and Australia, A/J mice do not have the mutated dysferlin gene, so researchers in those countries have imported the dysferlin deficient A/J strain (calling it A/J DYSF^PRMD) and use the A/J mice from their local mouse repositories as dysferlin sufficient controls.

C57BL/10 mice are the appropriate control for the C57BL/10.SJL-Dysf mouse strain, and C57BL/6 mice are the appropriate control for B6.A-Dysf^prmd/GeneJ mice, which contain the SJL and A/J mutations, respectively. The Jackson Laboratories recommend using wild type mice from their colony of B6.129-Dysf^tm1Kcam/J mice as controls for that targeted dysferlin deficient strain, but it is also important to note that this line has been backcrossed to C57BL/6 for seven generations, which should serve as an appropriate control strain.

The first Ferlin to be discovered was the protein Fer-1, in the nematode C. elegans. This protein is required for reproduction in C. elegans (it is involved in vesicle fusion during spermatid maturation), and therefore its name is an abbreviation for “fertilization factor 1”. The nomenclature for the human Ferlins, Fer1Lx, means “Fer-1 like number x”.

A deficiency of the Dysferlin protein causes the autosomal recessive disorders LGMD2B, and Miyoshi Myopathy. A lack of Otoferlin, which is normally expressed in the inner ear and brain, causes a recessive form of deafness, DFNB9. No human diseases have been associated with Myoferlin yet, although a mouse model lacking Myoferlin shows a myopathy and has very small muscles. The other three Ferlins, Fer1L4, Fer1L5, and Fer1L6, have not been associated with any diseases in humans or animals.

Although the genomes of most other mammals have not yet been analyzed as closely as the human genome, all of the identified Ferlins have also been identified in many other mammals: mice, rats, dogs, chimpanzees, cows, etc. The sequences are very similar to the human sequence. For instance, the human and mouse sequences for Dysferlin have 94% amino acid identity, and 97% amino acid similarity.