Gtp-bound Tubulin
Gtp-bound Tubulin - Related Videos
10:13
Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding
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Cited by 11 •
2017
Guanosine triphosphate (GTP) binding is one of the earliest events in G-Protein-Coupled Receptor (GPCR) activation. This protocol describes how to pharmacologically characterize specific GPCR-ligand interactions by monitoring the binding of the radio-labeled GTP analog, [35S]guanosine-5'-O-(3-thio)triphosphate ([35S]GTPγS), in response to a ligand of interest.
02:40
Second Harmonic Generation Imaging in a Rat Model to Study Tubulin Defects
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2025
Source: Piazza, V., et al., Label-Free Non-Linear Optics for the Study of Tubulin-Dependent Defects in Central Myelin. J. Vis. Exp. (2023)This video demonstrates the procedure for imaging microtubule abnormalities in a rat brain tissue slice using a two-photon excitation microscope. It employs second harmonic generation (SHG) signals to detect tubulin defects and reduced myelin production.
02:30
Isolation of Small Regulatory RNA–Bound Bacterial Target RNA Using Affinity Purification
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2025
Source: Mercier, N., et. al., MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria. J. Vis. Exp. (2021)This video demonstrates the isolation of MS2-tagged small regulatory RNA (sRNA) bound to its bacterial target RNA using affinity purification with a maltose-binding protein (MBP)–MS2 coat protein fusion immobilized on amylose resin.
09:42
Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides
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Cited by 6 •
2012
We describe here a technique that is now routinely used to isolate stably bound ribosome nascent chain complexes (RNCs). This technique takes advantage of the discovery that a 17 amino acid long SecM "arrest sequence" can halt translation elongation in a prokaryotic (E. coli) system, when inserted into (or fused to the C-terminus) of virtually any protein.
06:30
Optimizing Tubulin Yield from Porcine Brain Tissue
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2024
This protocol describes a technique for the high-yield isolation of tubulin from the porcine brain optimized for small-scale instrumentation. The isolation procedures are complemented by procedures for determining tubulin polymerization activity in vitro using co-sedimentation assays and transmission electron microscopy.
FAQs
GTP at the exchangeable site on β-tubulin favors a straight tubulin conformation. That structural state supports the addition of tubulin heterodimers at microtubule ends, making continued filament growth more likely. The nucleotide site therefore links the chemical state of an individual tubulin subunit to the physical behavior of the larger cytoskeletal polymer.
Once the tubulin is incorporated into the filament, GTP hydrolysis changes its contribution to microtubule stability. This transition can shift the filament from a growth-supporting state toward shrinkage. The sequence connects subunit incorporation, nucleotide conversion, and changes in filament behavior, allowing microtubules to reorganize rather than remain permanently assembled.
GTP hydrolysis provides a mechanism for switching microtubule stability after assembly. Tubulin carrying GTP promotes addition at growing ends, whereas the post-incorporation change can destabilize the filament and trigger shrinkage. This balance between extension and loss of subunits produces dynamic instability, a property that permits rapid cytoskeletal reorganization.
The two states support different filament behaviors. GTP-bound tubulin adopts the straight conformation associated with addition to growing microtubule ends, while hydrolysis after incorporation alters stability and may promote shrinkage. Comparing these states helps explain how the same protein building block can participate in both microtubule growth and disassembly.
Studies of this nucleotide-associated state can clarify how microtubules reorganize during cell division, intracellular transport, and changes in cell shape. Because these activities depend on controlled cytoskeletal rearrangement, examining the relationship between tubulin’s nucleotide state and filament stability helps connect molecular events with broader cellular behaviors.
GTP-bound tubulin provides a framework for studying how microtubule behavior responds to regulation. Its role in growth and subsequent stability changes is relevant to investigations of microtubule-targeting drugs, as well as cellular responses to mechanical or chemical signals. These applications use tubulin dynamics to examine how cells control cytoskeletal organization under changing conditions.