scholarly journals Effect of Phosphorylation in the Motor Domain of Human Myosin IIIA on Its ATP Hydrolysis Cycle

Biochemistry ◽  
2010 ◽  
Vol 49 (17) ◽  
pp. 3695-3702 ◽  
Author(s):  
Shigeru Komaba ◽  
Shinya Watanabe ◽  
Nobuhisa Umeki ◽  
Osamu Sato ◽  
Mitsuo Ikebe
Keyword(s):  
Atp Hydrolysis ◽  
Motor Domain ◽  
Myosin Iiia

scholarly journals Analysis of the Structural Mechanism of ATP Inhibition at the AAA1 Subunit of Cytoplasmic Dynein-1 Using a Chemical “Toolkit”

2021 ◽  
Vol 22 (14) ◽  
pp. 7704
Author(s):  
Sayi’Mone Tati ◽  
Laleh Alisaraie
Keyword(s):  
Binding Affinity ◽  
Atp Hydrolysis ◽  
Critical Role ◽  
Motor Protein ◽  
Structural Data ◽  
Retrograde Transport ◽  
Cytoplasmic Dynein ◽  
Motor Domain ◽  
Conformational Search ◽  
Structural Mechanism

Dynein is a ~1.2 MDa cytoskeletal motor protein that carries organelles via retrograde transport in eukaryotic cells. The motor protein belongs to the ATPase family of proteins associated with diverse cellular activities and plays a critical role in transporting cargoes to the minus end of the microtubules. The motor domain of dynein possesses a hexameric head, where ATP hydrolysis occurs. The presented work analyzes the structure–activity relationship (SAR) of dynapyrazole A and B, as well as ciliobrevin A and D, in their various protonated states and their 46 analogues for their binding in the AAA1 subunit, the leading ATP hydrolytic site of the motor domain. This study exploits in silico methods to look at the analogues’ effects on the functionally essential subsites of the motor domain of dynein 1, since no similar experimental structural data are available. Ciliobrevin and its analogues bind to the ATP motifs of the AAA1, namely, the walker-A (W-A) or P-loop, the walker-B (W-B), and the sensor I and II. Ciliobrevin A shows a better binding affinity than its D analogue. Although the double bond in ciliobrevin A and D was expected to decrease the ligand potency, they show a better affinity to the AAA1 binding site than dynapyrazole A and B, lacking the bond. In addition, protonation of the nitrogen atom in ciliobrevin A and D, as well as dynapyrazole A and B, at the N9 site of ciliobrevin and the N7 of the latter increased their binding affinity. Exploring ciliobrevin A geometrical configuration suggests the E isomer has a superior binding profile over the Z due to binding at the critical ATP motifs. Utilizing the refined structure of the motor domain obtained through protein conformational search in this study exhibits that Arg1852 of the yeast cytoplasmic dynein could involve in the “glutamate switch” mechanism in cytoplasmic dynein 1 in lieu of the conserved Asn in AAA+ protein family.

scholarly journals High-Resolution Crystal Structure and In Vivo Function of a Kinesin-2 Homologue in Giardia intestinalis

2008 ◽  
Vol 19 (7) ◽  
pp. 3124-3137 ◽  
Author(s):  
J. C. Hoeng ◽  
S. C. Dawson ◽  
S. A. House ◽  
M. S. Sagolla ◽  
J. K. Pham ◽  
...  
Keyword(s):  
High Resolution ◽  
Atp Hydrolysis ◽  
Intraflagellar Transport ◽  
Significant Inhibition ◽  
Giardia Intestinalis ◽  
Motor Domain ◽  
Parasitic Protist ◽  
Flagellar Assembly

A critical component of flagellar assembly, the kinesin-2 heterotrimeric complex powers the anterograde movement of proteinaceous rafts along the outer doublet of axonemes in intraflagellar transport (IFT). We present the first high-resolution structures of a kinesin-2 motor domain and an ATP hydrolysis–deficient motor domain mutant from the parasitic protist Giardia intestinalis. The high-resolution crystal structures of G. intestinalis wild-type kinesin-2 (GiKIN2a) motor domain, with its docked neck linker and the hydrolysis-deficient mutant GiKIN2aT104N were solved in a complex with ADP and Mg2+ at 1.6 and 1.8 Å resolutions, respectively. These high-resolution structures provide unique insight into the nucleotide coordination within the active site. G. intestinalis has eight flagella, and we demonstrate that both kinesin-2 homologues and IFT proteins localize to both cytoplasmic and membrane-bound regions of axonemes, with foci at cell body exit points and the distal flagellar tips. We demonstrate that the T104N mutation causes GiKIN2a to act as a rigor mutant in vitro. Overexpression of GiKIN2aT104N results in significant inhibition of flagellar assembly in the caudal, ventral, and posterolateral flagellar pairs. Thus we confirm the conserved evolutionary structure and functional role of kinesin-2 as the anterograde IFT motor in G. intestinalis.

Myosin Motor Domain Lever Arm Rotation Is Coupled to ATP Hydrolysis†

Biochemistry ◽  
2000 ◽  
Vol 39 (40) ◽  
pp. 12330-12335 ◽  
Author(s):  
Stefan Highsmith ◽  
Katherine Polosukhina ◽  
Don Eden
Keyword(s):  
Atp Hydrolysis ◽  
Motor Domain ◽  
Myosin Motor ◽  
Lever Arm ◽  
Myosin Motor Domain

scholarly journals Coupling of protein surface hydrophobicity change to ATP hydrolysis by myosin motor domain

1997 ◽  
Vol 72 (1) ◽  
pp. 18-23 ◽  
Author(s):  
M. Suzuki ◽  
J. Shigematsu ◽  
Y. Fukunishi ◽  
Y. Harada ◽  
T. Yanagida ◽  
...  
Keyword(s):  
Atp Hydrolysis ◽  
Surface Hydrophobicity ◽  
Motor Domain ◽  
Protein Surface ◽  
Myosin Motor ◽  
Myosin Motor Domain ◽  
Protein Surface Hydrophobicity

scholarly journals Elucidation of Structural Mechanism of ATP Inhibition at the AAA1 Subunit of Cytoplasmic Dynein 1 Using a Chemical "Toolkit"

2021 ◽  
Author(s):  
Sayi'Mone Martinet Tati ◽  
Laleh Alisaraie
Keyword(s):  
Binding Site ◽  
Binding Affinity ◽  
Atp Hydrolysis ◽  
Critical Role ◽  
Motor Protein ◽  
Structural Data ◽  
Retrograde Transport ◽  
Cytoplasmic Dynein ◽  
Motor Domain ◽  
Conformational Search

Dynein is a cytoskeletal motor protein that carries organelles via retrograde transport in eukaryotic cells. The motor protein belongs to the ATPase family of proteins associated with diverse cellular activities and plays a critical role in transporting cargoes to the minus end of the microtubules. The motor domain of dynein possesses a hexameric head, where ATP hydrolysis occurs. The AAA1 binding site is the leading ATP hydrolytic site, followed by the AAA3 subsite. Small-molecule ATP competitive inhibitors of dynein are thought to target the AAA1 site. The presented work elucidates the structure-activity relationship of dynapyrazole A and B, ciliobrevin A and D in their various protonated states and their 46 analogs for their binding properties in the nucleotide-binding site of the AAA1 subunit and their effects on the functionally essential subsites of the motor domain of cytoplasmic dynein 1, as there is currently no similar experimental structural data available. Ciliobrevin and its analogs bind to the ATP motifs of the AAA1, namely the Walker-A or P-loop, the Walker-B, and the sensor I and II. Ciliobrevin A shows a better binding affinity to the AAA1 binding site of dynein 1 than its D analog. Although the double bond in ciliobrevin A and D was expected to decrease the ligand potency, they show a better affinity to the AAA1 binding site than dynapyrazole A and B, lacking the bond. Protonation of the nitrogen in ciliobrevin A, D, dynapyrazole A, and B at the N9 site of ciliobrevin, and the N7 of the latter increased their binding affinity. Exploring ciliobrevin A geometrical configuration suggests the E isomer has a superior binding profile over the Z due to binding at the critical ATP motifs. Utilizing the refined structure of the motor domain obtained through protein conformational search in this study exhibits that Arg1852 of the yeast cytoplasmic dynein could involve in the "glutamate switch" mechanism in cytoplasmic dynein 1 in lieu of the conserved Asn in AAA+ protein family, as the guanidine moiety of the Arg engages in an H-bond with the carboxylate moiety of Glu1849.

scholarly journals The kinesin-5 tail domain directly modulates the mechanochemical cycle of the motor domain for anti-parallel microtubule sliding

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Tatyana Bodrug ◽  
Elizabeth M Wilson-Kubalek ◽  
Stanley Nithianantham ◽  
Alex F Thompson ◽  
April Alfieri ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Regulatory Mechanism ◽  
Atp Hydrolysis ◽  
Force Production ◽  
Bound State ◽  
Motor Domain ◽  
Tail Domain ◽  
Mitotic Spindles ◽  
Direct Binding ◽  
Mechanochemical Cycle

Kinesin-5 motors organize mitotic spindles by sliding apart microtubules. They are homotetramers with dimeric motor and tail domains at both ends of a bipolar minifilament. Here, we describe a regulatory mechanism involving direct binding between tail and motor domains and its fundamental role in microtubule sliding. Kinesin-5 tails decrease microtubule-stimulated ATP-hydrolysis by specifically engaging motor domains in the nucleotide-free or ADP states. Cryo-EM reveals that tail binding stabilizes an open motor domain ATP-active site. Full-length motors undergo slow motility and cluster together along microtubules, while tail-deleted motors exhibit rapid motility without clustering. The tail is critical for motors to zipper together two microtubules by generating substantial sliding forces. The tail is essential for mitotic spindle localization, which becomes severely reduced in tail-deleted motors. Our studies suggest a revised microtubule-sliding model, in which kinesin-5 tails stabilize motor domains in the microtubule-bound state by slowing ATP-binding, resulting in high-force production at both homotetramer ends.

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