Overview
Alternative Name M-TRTx-Gr1a, M-theraphotoxin-Gr1a, GsMTx4
Lyophilized Powder yes
Origin Grammostola rosea (Chilean rose) tarantula
Source Synthetic modified peptide
MW: 4435 Da
Form Lyophilized from double distilled water (ddH2O). May contain TFA as a residual counter ion.
Effective concentration 1.5 µM – 5 µM
Sequence GCLEFWWKCNPNDDKCCRPKLKCSKLFKLCNFSF-NH2
Modifications Disulfide bonds between: Cys2-Cys17, Cys9-Cys23, and Cys16-Cys30
Phe34 - C-terminal amidation
LC-Biotin
Phe34 - C-terminal amidation
LC-Biotin
Label LC-Biotin. The extent of labeling is 1 molecule of biotin per molecule of GsMTx-4.
Structure
Activity GsMTx-4 is an inhibitor of Na+ voltage-gated channels3 and cation-selective mechanosensitive channels1,2.
Shipping and storage The product is shipped as a lyophilized powder at room temperature. Upon receipt, store the product at -20°C. Protect from moisture.
Solubility Centrifuge the vial before adding solvent (10,000 x g for 5 minutes).
The lyophilizate may be difficult to visualize. Add solvent directly to the centrifuged vial. Tap the vial to aid in dissolving the lyophilized product. Tilt and gently roll the liquid over the walls of the vial. Avoid vigorous vortexing. Light vortexing for up to 3 seconds is acceptable if needed.
Soluble in pure water at high-micromolar concentrations (50 µM - 1 mM). For long-term storage in solution, it is recommended to prepare a stock solution by dissolving the product in double distilled water (ddH2O) at a concentration between 100-1000x of the final working concentration. Divide the stock solution into small aliquots and store at -20°C. Before use, thaw the relevant vial(s) and dilute to the desired working concentration in your working buffer. Centrifuge all product preparations at 10,000 x g for 5 minutes before use. It is recommended to prepare fresh solutions in working buffers just before use. Avoid multiple freeze-thaw cycles to maintain biological activity.
The lyophilizate may be difficult to visualize. Add solvent directly to the centrifuged vial. Tap the vial to aid in dissolving the lyophilized product. Tilt and gently roll the liquid over the walls of the vial. Avoid vigorous vortexing. Light vortexing for up to 3 seconds is acceptable if needed.
Soluble in pure water at high-micromolar concentrations (50 µM - 1 mM). For long-term storage in solution, it is recommended to prepare a stock solution by dissolving the product in double distilled water (ddH2O) at a concentration between 100-1000x of the final working concentration. Divide the stock solution into small aliquots and store at -20°C. Before use, thaw the relevant vial(s) and dilute to the desired working concentration in your working buffer. Centrifuge all product preparations at 10,000 x g for 5 minutes before use. It is recommended to prepare fresh solutions in working buffers just before use. Avoid multiple freeze-thaw cycles to maintain biological activity.
Storage of solutions Store the reconstituted solution at -20°C for the shortest time possible. Avoid multiple freeze-thaw cycles. We do not recommend storing the product in working solutions for longer than a day.
Our bioassay
Live cell imaging of GsMTx-4-Biotin in mouse M1 myeloid leukemia cells.(A-B) CellMask™ Actin 1X solution was applied for 30 minutes, resulting in an orange fluorescence to visualize cellular membrane. (C-D) Following this, the same cells underwent incubation with either 2.5 µM of GsMTx-4-Biotin (C) or GsMTx-4 (#STG-100) (D) for 45 minutes at 37ºC, followed by Alexa Streptavidin-647, leading to red fluorescence indicative of mechanosensitive ion channels distribution. (E-F) Live imaging of M1 cells allowed observation of GsMTx-4 distribution among the cells.
Expression of GsMTx-4-Biotin in Rat DRG Frozen Sections.(A-B) Anti-Piezo1 (extracellular) Antibody (#APC-087) was applied for 60 minutes at 37ºC, followed by staining with goat anti-rabbit AlexaFluor-488, resulting in green fluorescence predominantly in DRG cells. (C-D) The same frozen sections underwent incubation with either 2.5 µM of GsMTx-4-Biotin (C) or GsMTx-4 (#STG-100) (D) for 60 minutes at 37ºC. This was followed by staining with Alexa Streptavidin-647, resulting in red fluorescence in DRG cells. (E-F) Subsequently, CellMask™ Actin 1X solution was applied for 20 minutes to visualize the cellular membrane. (G-H) Co-staining of Anti-Piezo1 (extracellular) Antibody and GsMTx-4 in DRG cells is shown. In (G), the distribution of GsMTx-4 in mechanosensitive ion channels is indicated by arrows, whereas (H) does not show this distribution.
Direct flow cytometry of GsMTx-4 in live intact mouse M1 cells.___ M1 cells.
___ M1 cells + 1.5 µM GsMTx-4 (#STG-100) + 2 µg/ml Streptavidin, Alexa Fluor™ 647 conjugate.
___ M1 cells + 2 µg/ml Streptavidin, Alexa Fluor™ 647 conjugate.
___ M1 cells + 1.5 µM GsMTx-4-Biotin (#STG-100-B) + 2 µg/ml Streptavidin, Alexa Fluor™ 647 conjugate.
Alomone Labs GsMTx-4-Biotin inhibits NaV1.7 channel currents heterologously expressed in Xenopus oocytes.A. Representative time course of GsMTx-4-Biotin (#STG-100-B) inhibition of NaV1.7 channels current. Membrane potential was held at -100 mV, current was elicited by a 100 ms voltage step to 0 mV every 10 sec, and significantly inhibited by application of 0.5 μM GsMTx-4-Biotin (green).
B. Superimposed traces of NaV1.7 channel currents in the absence (control) and presence (green) of 0.5 μM GsMTx-4-Biotin (taken from the recording in A).
Scientific background
GsMTx-4 is a 34 amino acid peptidyl toxin originally isolated from the Grammostola rosea (Chilean rose) tarantula venom and belongs to the huwentoxin-1 family1.
This toxin inhibits different channels and in addition has antimicrobial activity. It blocks cation-selective mechanosensitive ion channels (strech-activated channels, SACs), without having an effect on whole-cell voltage-sensitive currents1. In addition, it inhibits atrial fibrillation2 as well as the membrane motor of outer hair cells3 at low doses. A medium toxicity on a large spectra of voltage-gated Na+ channels, namely NaV1.1, NaV1.2, NaV1.3, NaV1.4, NaV1.5, NaV1.6 and NaV1.7 was reported. GsMTx-4 also inhibits K+ channels KV11.1 and KV11.2, whereas it does not inhibit K+ channels KV1.1 (IC50 > 85 µM), KV1.4 (IC50 > 85 µM) and KV11.3 (IC50 = 53 µM)4. GsMTx-4 was also found to inhibit both TRPC1 and TRPC6 channels5,6, as well as Piezo1, the mechanosensitive channel7.
Antimicrobial activity is shown against several Gram-positive and Gram-negative bacteria as well8.
Target NaV1.7 channels, TRPC1, TRPC6, Piezo1
Lyophilized Powder
For research purposes only, not for human use
Last Update: 02/12/2024
Specifications
Citations
Citations
