Recombinant Poecilia reticulata Rhodopsin (rho)

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Cat. No.
BT2129156
Source
in vitro E.coli expression system
Synonyms
rho; Rhodopsin
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Description

Definition and Biological Role

Rhodopsin consists of an apoprotein (opsin) covalently bound to 11-cis-retinal, a vitamin A derivative . In Poecilia reticulata, this protein facilitates scotopic (low-light) vision. Recombinant versions are produced to investigate its biophysical properties, misfolding mechanisms (linked to retinal dystrophies), and therapeutic interventions .

Production Methods

Recombinant Poecilia reticulata rhodopsin is typically expressed in heterologous systems like E. coli or mammalian cells. A generalized protocol includes:

ParameterDetails
Expression SystemE. coli (common for high yield) or COS cells (for post-translational modifications) .
TagHis-tag for purification .
Protein LengthFull-length (e.g., 348 amino acids in pig rhodopsin) .
PurificationAffinity chromatography (Ni-NTA for His-tagged proteins) .
Reconstitution11-cis-retinal added in vitro to regenerate functional rhodopsin .

Disease Modeling

  • Retinitis Pigmentosa (RP): Class II mutations (e.g., P23H) cause ER retention and activate the unfolded protein response (UPR), leading to photoreceptor apoptosis . Recombinant mutants enable studies on proteostasis and therapeutic chaperones (e.g., YC-001) .

  • Constitutive Activity: Mutants like G90D exhibit light-independent signaling, modeling congenital stationary night blindness .

Therapeutic Development

  • Gene Therapy: AAV vectors deliver wild-type RHO to suppress dominant-negative mutants .

  • Pharmacological Chaperones: Compounds like 9-cis-retinal stabilize misfolded rhodopsin, improving trafficking to photoreceptor outer segments .

Challenges in Recombinant Production

  • Misfolding: Mutations disrupt disulfide bonding (e.g., Cys185–Cys187 in misfolded RP mutants), reducing retinal binding .

  • Low Stability: Misfolded rhodopsin degrades rapidly via the ubiquitin-proteasome system .

  • Species-Specific Variations: Post-translational modifications (e.g., glycosylation) differ across species, affecting functional studies .

Future Directions

  • High-Throughput Screening: Identify small molecules that rescue misfolded rhodopsin .

  • Cryo-EM Studies: Resolve Poecilia reticulata rhodopsin’s 3D structure to compare with mammalian orthologs .

  • Organoid Models: Use iPSC-derived retinal organoids to test gene therapies in vitro .

Product Specs

Form
Lyophilized powder
Note: While we prioritize shipping the format currently in stock, please specify your preferred format in order notes for customized preparation.
Lead Time
Delivery times vary depending on the purchasing method and location. Please contact your local distributor for precise delivery estimates.
Note: Shipping is standard with blue ice packs. Dry ice shipping requires prior arrangement and incurs additional charges.
Notes
Avoid repeated freeze-thaw cycles. Store working aliquots at 4°C for up to one week.
Reconstitution
Centrifuge the vial briefly before opening to collect the contents. Reconstitute the protein in sterile, deionized water to a concentration of 0.1-1.0 mg/mL. For long-term storage, we recommend adding 5-50% glycerol (final concentration) and aliquoting at -20°C/-80°C. Our standard glycerol concentration is 50%, which can be used as a reference.
Shelf Life
Shelf life depends on storage conditions, buffer composition, temperature, and protein stability. Generally, liquid formulations have a 6-month shelf life at -20°C/-80°C, while lyophilized formulations have a 12-month shelf life at -20°C/-80°C.
Storage Condition
Upon receipt, store at -20°C/-80°C. Aliquot to prevent repeated freeze-thaw cycles.
Tag Info
Tag type is determined during manufacturing.
The tag type is determined during production. If you require a specific tag, please inform us, and we will prioritize its development.
Synonyms
rho; Rhodopsin
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
1-354
Protein Length
full length protein
Species
Poecilia reticulata (Guppy) (Acanthophacelus reticulatus)
Target Names
rho
Target Protein Sequence
MNGTEGPYFYVPMVNTTGIVRSPYEYPQYYLVSPAAYACLGAYMFFLILVGFPINFLTLY VTIEHKKLRTPLNYILLNLAVADLFMVFGGFTTTIYTSMHGYFVLGRLGCNLEGYFATLG GEIGLWSLVVLAVERWLVVCKPISNFRFSENHAIMGLVFTWIMANSCAAPPLLGWSRYIP EGMQCSCGVDYYTRAEGFNNESFVIYMFICHFCIPLIVVFFCYGRLLCAVKEAAAAQQES ETTQRAEREVTRMVVIMVIGFLVCWIPYASVAWYIFTHQGSEFGPLFMTVPAFFAKSASI YNPLIYICMNKQFRHCMITTLCCGKNPFEEEEGASTTASKTEASSVSSSSVSPA
Uniprot No.
P79848

Target Background

Function
A photoreceptor essential for low-light vision. While most marine fish utilize retinal as a chromophore, freshwater species often employ 3-dehydroretinal, or a mixture of both. Light-induced isomerization of 11-cis to all-trans retinal triggers a conformational change, activating G-protein signaling. Subsequent receptor phosphorylation, mediated by arrestin, displaces the bound G-protein alpha subunit, terminating the signal.
Protein Families
G-protein coupled receptor 1 family, Opsin subfamily
Subcellular Location
Membrane; Multi-pass membrane protein. Cell projection, cilium, photoreceptor outer segment.

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