Recombinant Xenopus laevis Solute carrier family 25 member 46-A (slc25a46-a)

Molecular Structure

Recombinant SLC25A46-A from Xenopus laevis is a full-length protein comprising 417 amino acids (1-417aa) . The protein has a UniProt ID of Q6INQ6 and is commercially available with an N-terminal His-tag, expressed in E. coli expression systems . The complete amino acid sequence of the protein is as follows:

MQPRRPDRFDGLEYRGTSWGRGEGDPPPYQSSFPARSFSSSGDLSQHWVTTPPDIPGSRN LHWGDKSPQYGGADSNAGPPAFGEENSNSANSGEQLNRFAGFGIGLASLFTENVLAHPCI VLRRQCQVNYHARNYQLSPFNIVNIMYNFTKTQGLRALWKGMGSTFIVQGISLGAEGILS EFTHLPRELNHKWNPKQIGGHLLLKGLVYVIVTPFYSASLIETVQSEIIHDNPGILDCLK EGMGRVLNLGVPYSKRLLPLLVLTFPTVLHGILHYIVSSTVQKCVLFFIKKKSPPRLPAD GSNTVQNKLEDYFPELIANFAASLCADVLLYPLETVLHRLHIQGTRTIIDNTDLGHEVVP INTQYEGLKDCINTIKREEGGLGFYKGFGAVVVQYTLHAIVLQITKIIYSSVVQTSS

Physical and Biochemical Properties

The recombinant protein is typically supplied as a lyophilized powder with purity greater than 90% as determined by SDS-PAGE analysis . When reconstituted according to manufacturer specifications, it can be stored in appropriate buffer conditions for experimental use. The storage buffer typically consists of Tris/PBS-based buffer with 6% Trehalose at pH 8.0 .

Recombinant Expression

The production of Recombinant Xenopus laevis SLC25A46-A involves expression in E. coli bacterial systems, which allows for efficient production of the protein in its full-length form with an N-terminal His-tag . This expression system is widely used for recombinant protein production due to its scalability, cost-effectiveness, and ability to produce high yields of protein.

Comparative Function with Other SLC25 Family Members

Other members of the SLC25 family, such as SLC25A42, have been characterized as mitochondrial transporters for specific substrates. For instance, SLC25A42 has been shown to transport CoA and adenosine 3′,5′-diphosphate (PAP) with high specificity via a counter-exchange mechanism . While the specific transport substrates for SLC25A46 have not been definitively identified in the provided search results, its structural similarities to other SLC25 family members suggest it may also function as a specialized transporter within the mitochondrial membrane.

SLC25A46 in Bovine Models

Significant insights into SLC25A46 function have come from studying its role in bovine models. The "Turning calves syndrome," a sensorimotor polyneuropathy described in the French Rouge-des-Prés cattle breed, has been linked to a single nucleotide substitution in the SLC25A46 gene . This mutation causes a damaging amino acid substitution that affects the protein's function, leading to neurological symptoms in affected animals.

Mouse Knockout Models

To better understand SLC25A46 function, researchers have developed mouse knockout models. These studies revealed that disruption of the Slc25a46 gene affects not only the nervous system but also alters general metabolism, resulting in premature mortality . Specifically, Slc25a46 knockout mice exhibited:

1. Reduced growth rate despite normal initial feeding behavior

2. Ataxic gait, especially in the hind limbs, suggesting proprioception defects

3. Epileptic-like symptoms

4. Early mortality (between 3-4 weeks of life)

5. Multiple organ effects including smaller thymus, spleen, and liver

6. Biochemical abnormalities including increased biliary acids, bilirubin, and cholesterol

Neurological Disease Research

Given the clear links between SLC25A46 dysfunction and neurological symptoms in various animal models, recombinant Xenopus laevis SLC25A46-A provides a valuable tool for studying the molecular basis of neurodegenerative diseases. Researchers can use this protein to investigate:

1. Protein-protein interactions within the mitochondrial membrane

2. Structural studies to determine functional domains

3. In vitro assays to assess transport capabilities

4. Development of therapeutic approaches targeting mitochondrial dysfunction

Evolutionary and Comparative Studies

The availability of recombinant SLC25A46-A from Xenopus laevis, an amphibian model organism, enables comparative studies with mammalian counterparts. Such research can provide insights into the evolutionary conservation of mitochondrial dynamics mechanisms and how they may differ across vertebrate species.