Related Experiment Video
Updated: Jul 12, 2025
13:02
Production, Crystallization, and Structure Determination of the IKK-binding Domain of NEMO
Published on: December 28, 2019
7.5K
Solution NMR assignments and structure for the dimeric kinesin neck domain
Diana Seo1, Richard A Kammerer2, Andrei T Alexandrescu3
1Department of Molecular and Cell Biology, University of Connecticut, 91 N. Eagleville Road, Storrs, CT, 06269-3125, USA.
Biomolecular NMR Assignments
|October 20, 2023
Summary
The kinesin neck domain
Area of Science:
- Biochemistry
- Structural Biology
- Molecular Motors
Background:
- Kinesin is a motor protein essential for intracellular transport along microtubules.
- The kinesin heavy chain includes a neck domain connecting the motor head to cargo-binding regions.
- The neck domain's structure and flexibility are crucial for kinesin function.
Purpose of the Study:
- To determine the solution structure of the kinesin neck domain from rat Kif5c.
- To investigate the structural and dynamic properties of the neck domain and its linker region.
- To elucidate the role of the neck domain in kinesin's motor activity.
Main Methods:
- Nuclear Magnetic Resonance (NMR) spectroscopy (1H, 15N, 13C) was used for assignments and structure calculation.
- Analysis of Nuclear Overhauser Effect (NOE) data to determine interchain contacts in the dimer.
- Hydrogen-deuterium exchange experiments to assess solvent accessibility and stability.
Main Results:
- NMR assignments and a solution structure for the dimeric kinesin neck domain were obtained.
- The dimeric coiled coil structure is similar to the X-ray structure, while the linker region is disordered.
- The coiled coil is stable and protected from solvent, suggesting it does not unzip during kinesin walking.
Conclusions:
- The kinesin neck domain forms a stable dimeric coiled coil structure in solution.
- The disordered linker region acts as a flexible hinge connecting the motor head and neck.
- The stability of the coiled coil suggests alternative mechanisms for kinesin's motor function.
Related Concept Videos
01:15Overview of Myosin Structure and Function
4.4K
Myosins are a family of molecular motor proteins, first identified in the skeletal muscles, where they are responsible for muscle contraction. Along with their role in muscle contraction, these proteins also play a role in the intracellular transport of molecules and vesicles. There are twenty-four classes of myosins based on their domain sequence and organization. Of the twenty-four, six classes (Myosin I, Myosin II, Myosin V, Myosin VI, Myosin VII, and Myosin X) have been well...
4.4K
01:43The Movement of Organelles and Vesicles
4.5K
In eukaryotic cells, cytoskeletal filaments such as actin, microtubules, and intermediate filaments form a mesh-like cytoskeletal network. These filaments serve as tracks for transporting cellular cargo. Specialized motor proteins use the chemical energy stored in adenosine triphosphate (ATP) for this transport. During interphase, microtubules are polarized, with the plus-end towards the cell periphery and the minus-end towards the cell center. Two microtubule-associated motor proteins,...
4.5K
00:52¹H NMR of Conformationally Flexible Molecules: Temporal Resolution
868
At room temperature, the chair conformer of cyclohexane undergoes rapid ring flipping between two equivalent chair conformers at a rate of approximately 105 times per second. These two chair conformers are in equilibrium. The rapid ring flipping results in the interconversion of the axial proton to an equatorial proton and an equatorial to the axial proton. Such interconversions are too rapid and cannot be detected on the NMR timescale. Hence, the NMR spectrometer cannot distinguish between the...
868
01:18ATP Synthase: Structure
12.5K
ATP synthase or ATPase is among the most conserved proteins found in bacteria, mammals, and plants. This enzyme can catalyze a forward reaction in response to the electrochemical gradient, producing ATP from ADP and inorganic phosphate. ATP synthase can also work in a reverse direction by hydrolyzing ATP and generating an electrochemical gradient. Different forms of ATP synthases have evolved special features to meet the specific demands of the cell. Based on their specific feature, ATP...
12.5K
01:15¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR
1.1K
The axial and equatorial protons in cyclohexane can be distinguished by performing a variable-temperature NMR experiment. In this process, except for one proton, the remaining eleven protons are replaced by deuterium. The deuterium substitution avoids the possible peak splitting caused by the spin-spin coupling between the adjacent protons. The remaining proton flips between the axial and equatorial positions.
1.1K
01:13¹H NMR: Complex Splitting
1.3K
A proton M that is coupled to a proton X results in doublet signals for M. However, NMR-active nuclei can be simultaneously coupled to more than one nonequivalent nucleus. When M is coupled to a second proton A, such as in styrene oxide, each peak in the doublet is split into another doublet.
Splitting diagrams or splitting tree diagrams are routinely used to depict such complex couplings. While drawing splitting diagrams, the splitting with the larger coupling constant is usually applied...
Splitting diagrams or splitting tree diagrams are routinely used to depict such complex couplings. While drawing splitting diagrams, the splitting with the larger coupling constant is usually applied...
1.3K