Bistramide A

Protein target name:

Actin

Mechanism of action:

Severs actin filamentsCovalent sequestration of G-actin

Recommended Concentration for use in cells:

50-100 nM

Probe Information

Target and Probe Information

Target Details

Target class:

Structural protein

Subclass:

Cytoskeleton

HUGO ID:

HGNC:145

Entrez gene ID:

UNIPROT ID:

P63267

Target alias:

G-actin, F-actin

Probe Details

Chemical Probes Portal ID:

CPP738

Probe alias:

bistratene A

PubChem CID:

49864366

ChEMBL ID:

CHEMBL1221600

SMILES string:

C/C=C/C(=O)C[C@H]1CC[C@H](C)[C@H](CC(=O)NC[C@H](O)[C@H](C)C(=O)NCCC[C@H]2O[C@@]3(CCC[C@H](CC[C@H](C)/C=C(\C)[C@H](C)O)O3)CC[C@@H]2C)O1

InChi Key:

HXZRMADPDYFMEB-FTTMEYFSSA-N

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

7 nM

Potency assay:

Isothermal calorimetry (ITC) assays with G-actin and ATP. In G-actin polymerization experiments, Bistramide A decreased polymerization at concentrations 50-400 nM. In F-actin depolymerization assays, Bistramide A increased depolymerization at concentrations 120-450 nM. These results were confirmed in later experiments using TIRF microscopy (see PNAS paper).

PDB ID for probe-target interaction (3D structure):

2FXU

In cell validation

On target activity in cells

Potency assay (cells):

In A549 cell lysates, biotinylated Bistramide A interacted with a single protein that was shown to be beta-actin by MS. Covalently modified actin was detected in A549 cells treated with BODIPY-conjugated Bistramide A.

Target engagement assay (cells):

Yes, biotinylated Bistramide A interacted only with actin in A549 cells and BODIPY-conjugated Bistramide A bound covalently to monomeric actin.

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

Bistramide A has an IG50 ~1.5 uM (MTS assays) in multiple cell lines. It induces G2M arrest and polyploidy. It inhibits cytokinesis.

Primary Reference

Reference (doi):

10.1038/nchembio748 10.1021/ja058319c 10.1073/pnas.0710727105

Reference (PMID):

16372404 16551075 18334642

Endorsed probe

The Chemical Probes Portal endorses this compound as chemical probe for use in cells.

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

Portal staff comment: Bistramide A is not a PKC δ activator.

Jun 27 2017 - 7:46pm

Bistramide A is a novel actin-binding compound that disrupts actin filament formation. The compound has a fairly narrow concentration range and can be used as an orthogonal probe to study actin dynamics.

Jul 18 2017 - 11:44am

Bistramide A, at submicromolar concentrations, is shown to bind G-actin and sequester the actin molecule from polymerization into F-actin in cells, but at concentrations higher that 10 uM, the compound starts to inhibit PDBu binding to PKCdelta. So researchers should carefully set an experimental concentration range and should carefully interpret the consequences after treatment of cells with the compound, especially in growth inhibition or toxicity assays, which might yield phenotypes for multiple reasons.

Jul 28 2017 - 12:41pm

Bistramide A displays good affinity for actin with Kd=7nM. Its activity in cells is reported in the range of 20mM to 750nM (PMID 3238698 and 7807120) depending on the readout and cancer cell line tested. It is also active in normal human endothelial cells (PMID 3238698). Accordingly, a top probe concentration of 300-500nM might be used in parallel with the recommended concentration of 50-100 nM to cover all bases. This probe was tested in vivo but was reported to be too generally toxic to be able to study its anti-tumor activity (PMID 7807120); its testing in vivo is therefore not recommended.

Nov 16 2020 - 8:19pm

Bistramide A

Probe Information

Target Details

G-actin, F-actin

Probe Details

In vitro validation

In cell validation

Toxicity

Primary Reference

Supporting Organizations

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Bistramide A

Probe Information

Target Details

G-actin, F-actin

Probe Details

In vitro validation

In cell validation

Toxicity

Primary Reference

Supporting Organizations

Interested in supporting the portal?