Conformational Change of the Loop L5 in Rice Kinesin Motor Domain Induced by Nucleotide Binding

2006 ◽  
Vol 139 (5) ◽  
pp. 857-864 ◽  
Author(s):  
Nobuhisa Umeki ◽  
Toshiaki Mitsui ◽  
Kazunori Kondo ◽  
Shinsaku Maruta
Keyword(s):  
Conformational Change ◽  
Nucleotide Binding ◽  
Motor Domain ◽  
Kinesin Motor

scholarly journals The divergent mitotic kinesin MKLP2 exhibits atypical structure and mechanochemistry

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Joseph Atherton ◽  
I-Mei Yu ◽  
Alexander Cook ◽  
Joseph M Muretta ◽  
Agnel Joseph ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Motor Function ◽  
Nucleotide Binding ◽  
Motor Domain ◽  
Kinesin Motor ◽  
Binding Motifs ◽  
Cryo Electron Microscopy ◽  
Specific Sequences

MKLP2, a kinesin-6, has critical roles during the metaphase-anaphase transition and cytokinesis. Its motor domain contains conserved nucleotide binding motifs, but is divergent in sequence (~35% identity) and size (~40% larger) compared to other kinesins. Using cryo-electron microscopy and biophysical assays, we have undertaken a mechanochemical dissection of the microtubule-bound MKLP2 motor domain during its ATPase cycle, and show that many facets of its mechanism are distinct from other kinesins. While the MKLP2 neck-linker is directed towards the microtubule plus-end in an ATP-like state, it does not fully dock along the motor domain. Furthermore, the footprint of the MKLP2 motor domain on the MT surface is altered compared to motile kinesins, and enhanced by kinesin-6-specific sequences. The conformation of the highly extended loop6 insertion characteristic of kinesin-6s is nucleotide-independent and does not contact the MT surface. Our results emphasize the role of family-specific insertions in modulating kinesin motor function.

scholarly journals Structure and comparison of the motor domain of centromere-associated protein E

2021 ◽  
Vol 77 (3) ◽  
pp. 280-287
Author(s):  
Asuka Shibuya ◽  
Naohisa Ogo ◽  
Jun-ichi Sawada ◽  
Akira Asai ◽  
Hideshi Yokoyama
Keyword(s):  
Binding Site ◽  
Anticancer Drugs ◽  
Structural Information ◽  
Nucleotide Binding ◽  
Competitive Inhibitor ◽  
Nucleotide Binding Site ◽  
Motor Domain ◽  
Kinesin Motor ◽  
Protein Preparation ◽  
Rate Limiting Step

Centromere-associated protein E (CENP-E) plays an essential role in mitosis and is a target candidate for anticancer drugs. However, it is difficult to design small-molecule inhibitors of CENP-E kinesin motor ATPase activity owing to a lack of structural information on the CENP-E motor domain in complex with its inhibitors. Here, the CENP-E motor domain was crystallized in the presence of an ATP-competitive inhibitor and the crystal structure was determined at 1.9 Å resolution. In the determined structure, ADP was observed instead of the inhibitor in the nucleotide-binding site, even though no ADP was added during protein preparation. Structural comparison with the structures of previously reported CENP-E and those of other kinesins indicates that the determined structure is nearly identical except for several loop regions. However, the retention of ADP in the nucleotide-binding site of the structure strengthens the biochemical view that the release of ADP is a rate-limiting step in the ATPase cycle of CENP-E. These results will contribute to the development of anticancer drugs targeting CENP-E and to understanding the function of kinesin motor domains.

scholarly journals Activation mechanism of ATP-sensitive K+ channels explored with real-time nucleotide binding

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Michael Puljung ◽  
Natascia Vedovato ◽  
Samuel Usher ◽  
Frances Ashcroft
Keyword(s):  
Conformational Change ◽  
Ionic Currents ◽  
Nucleotide Binding ◽  
Activation Mechanism ◽  
Time Resolved ◽  
K Channels ◽  
Novel Approach ◽  
Binding Event ◽  
Fluorescent Nucleotides

The response of ATP-sensitive K+ channels (KATP) to cellular metabolism is coordinated by three classes of nucleotide binding site (NBS). We used a novel approach involving labeling of intact channels in a native, membrane environment with a non-canonical fluorescent amino acid and measurement (using FRET with fluorescent nucleotides) of steady-state and time-resolved nucleotide binding to dissect the role of NBS2 of the accessory SUR1 subunit of KATP in channel gating. Binding to NBS2 was Mg2+-independent, but Mg2+ was required to trigger a conformational change in SUR1. Mutation of a lysine (K1384A) in NBS2 that coordinates bound nucleotides increased the EC50 for trinitrophenyl-ADP binding to NBS2, but only in the presence of Mg2+, indicating that this mutation disrupts the ligand-induced conformational change. Comparison of nucleotide-binding with ionic currents suggests a model in which each nucleotide binding event to NBS2 of SUR1 is independent and promotes KATP activation by the same amount.

scholarly journals The mechanism of kinesin inhibition by kinesin-binding protein

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Joseph Atherton ◽  
Jessica JA Hummel ◽  
Natacha Olieric ◽  
Julia Locke ◽  
Alejandro Peña ◽  
...  
Keyword(s):  
Cell Function ◽  
Binding Protein ◽  
Eukaryotic Cell ◽  
Motor Domain ◽  
Tetratricopeptide Repeat ◽  
Kinesin Motor ◽  
Temporal Regulation ◽  
Cryo Electron Microscopy ◽  
Local Environments ◽  
Binding Surface

Subcellular compartmentalisation is necessary for eukaryotic cell function. Spatial and temporal regulation of kinesin activity is essential for building these local environments via control of intracellular cargo distribution. Kinesin-binding protein (KBP) interacts with a subset of kinesins via their motor domains, inhibits their microtubule (MT) attachment, and blocks their cellular function. However, its mechanisms of inhibition and selectivity have been unclear. Here we use cryo-electron microscopy to reveal the structure of KBP and of a KBP–kinesin motor domain complex. KBP is a tetratricopeptide repeat-containing, right-handed α-solenoid that sequesters the kinesin motor domain’s tubulin-binding surface, structurally distorting the motor domain and sterically blocking its MT attachment. KBP uses its α-solenoid concave face and edge loops to bind the kinesin motor domain, and selected structure-guided mutations disrupt KBP inhibition of kinesin transport in cells. The KBP-interacting motor domain surface contains motifs exclusively conserved in KBP-interacting kinesins, suggesting a basis for kinesin selectivity.

scholarly journals Nucleotide binding triggers a conformational change of the CBS module of the magnesium transporter CNNM2 from a twisted towards a flat structure

2014 ◽  
Vol 464 (1) ◽  
pp. 23-34 ◽  
Author(s):  
María Ángeles Corral-Rodríguez ◽  
Marchel Stuiver ◽  
Guillermo Abascal-Palacios ◽  
Tammo Diercks ◽  
Iker Oyenarte ◽  
...  
Keyword(s):  
Conformational Change ◽  
Nucleotide Binding ◽  
Conformational Flexibility ◽  
Flat Structure ◽  
Magnesium Transporter

Nucleotide binding triggers a conformational change of the Bateman module of the magnesium transporter CNNM2. The hypomagnesaemia-causing mutation T568I impairs MgATP binding and limits the conformational flexibility of this protein module.

Kinetic Mechanism of Kinesin Motor Domain

Biochemistry ◽  
1995 ◽  
Vol 34 (40) ◽  
pp. 13233-13241 ◽  
Author(s):  
Y. Z. Ma ◽  
Edwin W. Taylor
Keyword(s):  
Kinetic Mechanism ◽  
Motor Domain ◽  
Kinesin Motor

scholarly journals The family-specific K-loop influences the microtubule on-rate but not the superprocessivity of kinesin-3 motors

2014 ◽  
Vol 25 (14) ◽  
pp. 2161-2170 ◽  
Author(s):  
Virupakshi Soppina ◽  
Kristen J. Verhey
Keyword(s):  
Bound State ◽  
Motor Domain ◽  
Cellular Transport ◽  
Kinesin Motor ◽  
Cargo Transport ◽  
The Family ◽  
Initial Interaction ◽  
Kinesin Superfamily ◽  
Positively Charged ◽  
Functional Output

The kinesin-3 family (KIF) is one of the largest among the kinesin superfamily and an important driver of a variety of cellular transport events. Whereas all kinesins contain the highly conserved kinesin motor domain, different families have evolved unique motor features that enable different mechanical and functional outputs. A defining feature of kinesin-3 motors is the presence of a positively charged insert, the K-loop, in loop 12 of their motor domains. However, the mechanical and functional output of the K-loop with respect to processive motility of dimeric kinesin-3 motors is unknown. We find that, surprisingly, the K-loop plays no role in generating the superprocessive motion of dimeric kinesin-3 motors (KIF1, KIF13, and KIF16). Instead, we find that the K-loop provides kinesin-3 motors with a high microtubule affinity in the motor's ADP-bound state, a state that for other kinesins binds only weakly to the microtubule surface. A high microtubule affinity results in a high landing rate of processive kinesin-3 motors on the microtubule surface. We propose that the family-specific K-loop contributes to efficient kinesin-3 cargo transport by enhancing the initial interaction of dimeric motors with the microtubule track.

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