Functional adaptation of the switch-2 nucleotide sensor enables rapid processive translocation by myosin-5

Nikolett T. Nagy, Takeshi Sakamoto, Balázs Takács, Máté Gyimesi, Eszter Hazai, Zsolt Bikádi, James R. Sellers, M. Kovács

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Active site loops that are conserved across superfamilies of myosins, kinesins, and G proteins play key roles in allosteric coupling of NTP hydrolysis to interaction with track filaments or effector proteins. In this study, we investigated how the class-specific natural variation in the switch-2 active site loop contributes to the motor function of the intracellular transporter myosin-5. We used single-molecule, rapid kinetic and spectroscopic experiments and semiempirical quantum chemical simulations to show that the class-specific switch-2 structure including a tyrosine (Y439) in myosin-5 enables rapid processive translocation along actin filaments by facilitating Mg 2+-dependent ADP release. Using wild-type control and Y439 point mutant myosin-5 proteins, we demonstrate that the translocation speed precisely correlates with the kinetics of nucleotide exchange. Switch-2 variants can thus be used to fine-tune translocation speed while maintaining high processivity. The class-specific variation of switch-2 in various NTPase superfamilies indicates its general role in the kinetic tuning of Mg2+-dependent nucleotide exchange.

Original languageEnglish
Pages (from-to)4480-4490
Number of pages11
JournalFASEB Journal
Volume24
Issue number11
DOIs
Publication statusPublished - Nov 2010

Fingerprint

Myosins
Nucleotides
Switches
Sensors
Kinetics
Catalytic Domain
Nucleoside-Triphosphatase
Kinesin
Protein Transport
Actin Cytoskeleton
GTP-Binding Proteins
Adenosine Diphosphate
Tyrosine
Actins
Hydrolysis
Proteins
Tuning
Molecules
Experiments

Keywords

  • Actomyosin
  • Kinetics
  • Nucleotide exchange
  • Single molecule
  • Sliding speed

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics
  • Molecular Biology

Cite this

Functional adaptation of the switch-2 nucleotide sensor enables rapid processive translocation by myosin-5. / Nagy, Nikolett T.; Sakamoto, Takeshi; Takács, Balázs; Gyimesi, Máté; Hazai, Eszter; Bikádi, Zsolt; Sellers, James R.; Kovács, M.

In: FASEB Journal, Vol. 24, No. 11, 11.2010, p. 4480-4490.

Research output: Contribution to journalArticle

Nagy, NT, Sakamoto, T, Takács, B, Gyimesi, M, Hazai, E, Bikádi, Z, Sellers, JR & Kovács, M 2010, 'Functional adaptation of the switch-2 nucleotide sensor enables rapid processive translocation by myosin-5', FASEB Journal, vol. 24, no. 11, pp. 4480-4490. https://doi.org/10.1096/fj.10-163998
Nagy, Nikolett T. ; Sakamoto, Takeshi ; Takács, Balázs ; Gyimesi, Máté ; Hazai, Eszter ; Bikádi, Zsolt ; Sellers, James R. ; Kovács, M. / Functional adaptation of the switch-2 nucleotide sensor enables rapid processive translocation by myosin-5. In: FASEB Journal. 2010 ; Vol. 24, No. 11. pp. 4480-4490.
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