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CNST

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All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

Background

CNST, also known as consortin or connexin sorting protein is a 673-amino acid single-pass transmembrane protein with an estimated size of 70 kDa coded by the human chromosomal location 1q44. CNST has ubiquitous expression in human tissues, with high (RPKM 10.5), moderate (brain: RPKM 9.5) and low levels of RNA across all other tested tissue types from the same individuals, exceeding genome-wide average values through a total of at least 25 human normal adult/male-tissue comparisons consistent with its universal significance in cellular protein trafficking. Localization studies at the subcellular level revealed that CNST is enriched in tubulovesicular transport organelles, mostly found within the trans-Golgi network (TGN), and at or near plasma membrane sites of anterograde trafficking to cell surface locations where it serves as a cargo receptor for TMD-containing proteins exported from the Golgi complex. Originally discovered as a connexin binding partner, CNST was later identified to be the first TGN cargo receptor with specificity in targeting transmembrane proteins towards the plasma membrane. This interacts directly with an essential group of clathrin adaptors, the GGA1 and 2 proteins that serve to connect connexin-containing vesicles placed in a secure point between AP-1/clathrin coat machinery involved in regulating transport from the TGN to plasma membrane. In addition to its canonical role in connexin trafficking, burgeoning evidence has also implicated CNST in various disease processes including acute myeloid leukemia (AML), where increased levels of expression are representative of a leukemic stem cell phenotype and correlate with an unfavorable clinical outcome; inherited non-syndromic hearing loss (DFNB1); regulation of bone mass deposition; as well as depolarization from TGF-β1 stimulation-induced cardiac fibroblasts.

Fig.1 CNST expression is associated with impaired hematopoietic differentiation. (OA Literature)Fig.1 CNST expression correlates with poor differentiation of the hematopoietic system.1

CNST Protein Function: A Master Coordinator of Gap Junction Dynamics

Because of its versatile molecular interactions, the functional repertoire of CNST encompasses membrane protein trafficking, gap junction assembly and cellular communication:

  • Trans-Golgi Network Cargo Receptor Function: CNST It binds connexins directly through its intrinsically disordered N-terminal region in addition to simultaneously interacting with GGA1 and GGA2 clathrin adaptors via the DXXLL motif. This dual-binding ability allows CNST to identify connexin cargo and recruit the clathrin coat machinery, which promotes vesicle budding from TGN leading its traffic towards cell surface. Mutagenesis of the DXXLL motif disrupts its interaction with AP2 and leads to intracellular accumulation of connexins, indicating that this unique interaction is required for efficient delivery to the plasma membrane.
  • Connexin Recycling and Gap Junction Plaque Degradation: CNST has a dual role in connexin homeostasis by regulating both anterograde trafficking and endocytic recycling. Inhibition of CNST by RNA interference impedes both matrix targeting and subsequent processing to the cell surface so that connexins accumulate intracellularly. By contrast, depletion and re-distribution of the CNST pool postpone gap junction plaque degradation in cells indicating that CNST may also mediate recycling of internalised connexins to plasma membrane or routing towards degradation pathways. This bidirectional trafficking control provides a central function for CNST in regulating both gap junction half-life and intercellular communication signal strength.
  • Connexin Subtype Specificity and Broad Cargo Recognition: CNST interacts with Cx43, the most abundant intercellular channel protein in diverse cell types; multiple other connexins including Cx32 (myocytes), Cx26 (keratinocyte and myotube mutations); and Cx50 (lens epithelial cells). And because the N-terminal region is intrinsically disordered, it could be able to accommodate relatively promiscuous connexin interaction interfaces for various gap junction proteins and thus CNST may act as a generic sorting receptor of bespoken ganglionic connection. Additionally, CNST may act to select additional transmembrane cargo proteins or unrelated membrane protein structures to which it can bind through increasing structural similarity with the connexins and shared sorting signals.

CNST Membrane Protein Product

Creative Biolabs provides a flexible range of high-quality CNST membrane protein products to support structural and functional studies of this trans-Golgi network cargo receptor. Our membrane protein platform can provide CNST constructs and preparations suitable for a range of research applications, including antibody development and characterization, binding assays, GGA1/GGA2 interaction studies, and investigations of connexin trafficking and TGN sorting mechanisms. These products can serve as research tools for studying CNST interactions with connexins, clathrin adaptors, and related trafficking machinery. Available formats and construct designs may vary according to specific experimental requirements.

CNST Protein Product

Not finding the membrane protein product you need? Contact us to start your one-stop custom service!

CNST Stable Cell Line Product

Establishing stable cellular models is essential in the CNST field. Designed from the ground up, Creative Biolabs provides custom-engineered CNST membrane protein stable cell lines working with target CNST-expressing cells at high levels consistently. These stable cell lines would be suitable for screening assays, connexin trafficking assays, studies of gap junction plaque formation / turnover and endocytic recycling as well high-throughput screening providing a reproducible tool-kit by which to probe CNST biology & pharmacology. We help you get the most expression with healthy cells, which drives efficiency in your research pipeline. This includes CNST-overexpressing lines for studying connexin targeting and GGA adaptor recruitment, as well as CNST-knockdown lines to study loss of function; expression is validated by flow cytometry (top), Western blotting (bottom left) or functional CCX surface delivery assays (lower right). Connexin-GFP-fusion based specialized-reporter cell lines allow real-time imaging of gap junction plaque dynamics, establishing important tools for high-throughput-mechanistic-dissection-screening in cardiac ischemia and neuropathy models.

CNST Stable Cell Line Product

Not finding the stable cell line product you need? Contact us to start your one-stop custom service!

CNST Recombinant Antibody Product

We are at the forefront of producing high-affinity recombinant antibodies targeting CNST tailored to meet the most challenging and technically demanding requirements of your research applications. These recombinant antibodies are more specific, sensitive and consistent from batch-to-batch compared to conventional polyclonal. Our CNST recombinant antibodies are tested for WB, ELISA, FCM, IF (Immunofluorescence), ICC (Immunocytochemistry), IHC(Immunohistochemistry) and IP(Immunoprecipitation) validation to enable reliable detection of the target protein from cardiac tissue homogenates; cochlear extracts; tumor biopsies; cell lysate. We provide antibodies against unique CNST epitopes, including the N-terminal intrinsically disordered region for interaction studies with connexins, the DXXLL motif for GGA adaptor binding sites identification and mapping in cellular context, a transmembrane domain to assess membrane integration potentials, C terminal tail domains which enables localisation between target organelles.

CNST Recombinant Antibody Product

Not finding the recombinant antibody product you need? Contact us to start your one-stop custom service!

Product Features

  • High Purity & Activity: High purity and verified integrity for dependable downstream work.
  • Diverse Expression Systems: Choice of expression formats to match routine or specialized projects.
  • Species Reactivity: Products available for human, mouse, pig, and other species.
  • Validated Applications: Application-tested to deliver consistent signals across common lab workflows.
  • Superior Batch-to-Batch Consistency: Stringent quality controls to ensure low variability from batch to batch.
  • Customization Options: Tags/Labels/package Modifications available upon request.

Custom CNST Membrane Protein and Antibody Discovery Services

Apart from our catalog products, Creative Biolabs also provides specific custom services for membrane protein and antibody discovery & development. With the extensive experience and best-in-class platforms we offer, specifically focus on helping you in:

  • Production & Validation: Cloning of the synthesized optimized gene into an expression vector, plus small-scale expression trials to help determine the best expression conditions.
  • Custom Antibody Production: Produced via large-scale expression in mammalian/insect/cell culture or bacterial expression systems, purified in detergent or reconstituted into nanodiscs.
  • Engineered Cell Lines: Creation of stable cell lines that physiologically mimic endogenous CNST expression and activity.
  • Functional Assessment Systems: Designing and executing assays to assess protein function and ligand binding.

Frequently Asked Questions (FAQ)

  1. Are Creative Biolabs'CNST products suitable for clinical or diagnostic use?

    No. All Creative Biolabs CNST products and services are for research use only, not clinical diagnosis or therapy of any disease(s) prevention treatment or cure.

  2. Can Creative Biolabs make CNST copies to study disease's methods of transmission?

    Absolutely. Our custom protein engineering services accommodate a wide range of modifications. Our scientific team collaborates closely with clients to design constructs optimized for trafficking studies, structural biology, GGA adaptor interaction mapping, and screening research applications.

  3. Do you provide antibodies that detect the CNST as it assembles into a connexin-bound, trafficking-competent complex at the ER membrane?

    Yes, we offer conformation-dependent antibodies that recognize CNST conformational epitopes exposed by the binding of either cx26 or cx30 C-terminal. These reagents allow for co-immunoprecipitation and immunofluorescence visualization of the intact ER-resident assembly complex in fixed cells or membrane preparations while generating minimal signal against unpartnered monomeric species dispersed throughout the cytosol.

  4. Do you provide stable cell lines that co-expresses CNST with connexin-26 or Cx30 to model continuous gap junction biogenesis, plaque assembly kinetics and intercellular communication restoration?

    Yes, we have created pairs of dual-stable lines where both full-length CNST and a pair of selected connexin partners are constitutively expressed from independent loci. Each batch comes with a documented gap junction plaque density (GJPD) and dye-coupling efficiency, which allows to use well-annotated standardized cellular platform for continuous mechanistic studies without the need of transient transfection.

Reference
  1. Liu, Haoyu, et al. "CNST is characteristic of leukemia stem cells and is associated with poor prognosis in AML." Frontiers in Pharmacology 13 (2022): 888243. Under Open Access license CC BY 4.0, without modification. https://doi.org/10.3389/fphar.2022.888243
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