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Investigating MAPs and Tubulin?

Microtubule Associated Proteins (MAPs) are proteins that interact with tubulin and microtubules to regulate their function and also to transport cargo. The well known MAPs MAP1 & 2 and Tau are expressed in CNS material and help stabilize microtubule structures. Cytoskeleton's BK029 kit is an effective and efficient tool for identification of novel and established MAPs.

Microtubule Binding Protein Spin-Down Assay Biochem Kit

(Cat. # BK029) Product Uses:

·     To determine whether a protein or compound binds to microtubules.

·     To test various deletion mutants for their ability to still bind microtubules, then calculate their affinity for microtubules, and hence identify the microtubule binding site.

·     In the presence of a non-hydrolyzable analog of ATP (kinesins) or GTP (dynamin) extract these from cells or show their binding affinity when mutated.

Above: MAP Spin-Down Assay using BK029. Microtubules were mixed with buffer (MT), MAP proteins (MT + MAPF) or BSA (MT +BSA) and MTs were pelleted by centrifugation at 100,000 x g. Supernatant (S) and Pellet (P) fractions were examined by SDS-PAGE. MAP proteins, but not BSA, bind to MTs and co-precipitate. MAP (MAPF) or BSA (BSA) proteins alone do not pellet.

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Recent Citations for BK029

 

Hori T. et al. Microtubule assembly by tau impairs endocytosis and neurotransmission via dynamin sequestration in Alzheimer's disease synapse model. Elife. 2022 Apr 26;11:e73542. doi: 10.7554/eLife.73542.

 

Wortzel I. et al. Mitotic HOOK3 phosphorylation by ERK1c drives microtubule-dependent Golgi destabilization and fragmentation. iScience. 2021 May 31;24(6):102670. doi: 10.1016/j.isci.2021.102670.

This brief review will focuses on tubulin and colchicine as a possible target as a covid therapy.

Alternative Tools For Studying Tubulin and MAPs

Comparison of SPY-labeled HeLa cells imaged with confocal vs. STED microscopy. Imaged with 93X objective and provided courtesy of Spirochrome.

Spirochrome Tubulin Imaging Tools:

SiR-Tubulin Kit (CY-SC002)

Cytoskeleton Kit (SiR Actin and Tubulin) (CY-SC006)

SPY555-Tubulin Kit (CY-SC203)

SPY650-Tubulin Kit (CY-SC503)

Select MAP proteins available from Cytoskeleton Inc.

Cytoskeleton purified MAPs:

Eg5 Kinesin Motor Domain Protein (EG01)

CENP-E Kinesin Motor Domain Protein (CP01)

BimC Kinesin Motor Domain Protein (BM01)