Recombinant Mouse Palmitoyltransferase ZDHHC7 (Zdhhc7)
Description
Functional Roles
ZDHHC7 catalyzes S-palmitoylation, transferring palmitate groups to cysteine residues on target proteins. Key findings include:
Mechanistic Insights
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Substrate Specificity: Directly palmitoylates NLRP3 at Cys126, promoting inflammasome assembly and ASC oligomerization .
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Subcellular Localization: Mediates NLRP3 trafficking to the trans-Golgi network (TGN) in resting macrophages and dispersed TGN during activation .
Inflammasome Studies
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Knockout Models: Zdhhc7-null macrophages show reduced NLRP3 activation and IL-1β secretion .
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Therapeutic Potential: Targeting ZDHHC7 suppresses NLRP3-driven inflammation in murine peritonitis models .
Chemical-Proteomic Profiling
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Substrate Mapping: Engineered ZDHHC7 mutants paired with "bumped" lipid probes enable proteome-wide identification of S-acylated proteins .
Technical Considerations
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Storage: Lyophilized powder stable at -80°C; reconstitute in Tris/PBS buffer with 6% trehalose .
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Activity Assays: Validated via fluorescence resonance energy transfer (FRET) and co-immunoprecipitation (co-IP) .
Comparative Analysis with Human Orthologs
Product Specs
Note: We will prioritize shipping the format currently in stock. However, if you have specific requirements for the format, please indicate them in your order. We will prepare the product according to your specifications.
Note: All of our proteins are shipped with standard blue ice packs. If you require dry ice shipping, please communicate with us in advance as additional fees will apply.
Generally, the shelf life of the liquid form is 6 months at -20°C/-80°C. The shelf life of the lyophilized form is 12 months at -20°C/-80°C.
Target Background
- DHHC7 knockout mice developed hyperglycemia and glucose intolerance, confirming that DHHC7 is the primary PAT for Glut4 and that this mechanism is essential for insulin-regulated glucose homeostasis. PMID: 28057756
Q&A
Basic Research Questions
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What is ZDHHC7 and what is its primary function in cellular systems?
ZDHHC7 is a member of the DHHC domain-containing protein family that functions as a palmitoyltransferase enzyme. Its primary role is catalyzing the addition of palmitate, a 16-carbon fatty acid, to specific cysteine residues in target proteins through a thioester bond (S-palmitoylation). This post-translational modification is critical for regulating protein localization, stability, and function.
ZDHHC7 is expressed in various tissues and cell types, with notable expression patterns in prostate epithelial cells. Studies have demonstrated that ZDHHC7 expression is frequently decreased in cancer cell lines compared to normal epithelial cells, suggesting potential tumor suppressor functions .
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What are the validated substrates of mouse ZDHHC7?
ZDHHC7 has several confirmed substrates involved in cellular polarity and signaling:
Researchers have established that ZDHHC7-mediated palmitoylation of Scribble is particularly critical for maintaining proper cellular polarity. When ZDHHC7 expression is lost or reduced, Scribble palmitoylation decreases, leading to its mislocalization from cell membranes .
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What methods are used to detect and measure ZDHHC7-mediated protein palmitoylation?
Several complementary approaches can be employed to detect ZDHHC7-mediated palmitoylation:
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Acyl-Biotin Exchange (ABE) Assay: This method allows for the detection of palmitoylated proteins by replacing palmitate groups with biotin, followed by pull-down with streptavidin and western blotting.
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Metabolic Labeling: Cells are treated with alkyne or azide-modified palmitate analogs that can be conjugated to reporter molecules via click chemistry.
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Palmitoylation-specific Antibodies: For detection of specific palmitoylated proteins when available.
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Confocal Microscopy: To visualize the localization of potential substrates like Scribble, which demonstrates membrane localization when properly palmitoylated .
Researchers commonly validate palmitoylation by expressing wild-type proteins alongside palmitoylation-deficient mutants (e.g., SCRIB C4/10S mutant) in cell lines to compare cellular localization and function .
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How can researchers generate functional recombinant mouse ZDHHC7 for in vitro studies?
Producing functional recombinant ZDHHC7 requires careful attention to its transmembrane domain structure:
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Expression System Selection: Mammalian expression systems (HEK293 cells) are preferred over bacterial systems to ensure proper folding and post-translational modifications.
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Vector Design: Incorporate appropriate tags (His, FLAG, or GST) at the N-terminus to avoid disrupting the catalytic DHHC domain.
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Solubilization Strategy: Use mild detergents (DDM or CHAPS) during membrane protein extraction to maintain enzymatic activity.
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Activity Verification: Confirm enzyme activity through in vitro palmitoylation assays using known substrates like Scribble.
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Storage Considerations: Store with glycerol at -80°C with protease inhibitors to maintain stability and enzymatic activity.
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