The kinase domain alters the kinetic properties of the myosin IIIA motor

Andréa C. Dosé, Shobana Ananthanarayanan, Judy E. Moore, Amoreena C. Corsa, Beth Burnside, Christopher Yengo

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Myosin IIIA is unique among myosin proteins in that it contains an N-terminal kinase domain capable of autophosphorylating sites on the motor domain. A construct of myosin IIIA lacking the kinase domain localizes more efficiently to the stereocilia tips and alters the morphology of the tips in inner ear hair cells. Therefore, we performed a kinetic analysis of myosin IIIA without the kinase domain (MIII ΔK) and compared these results with our reported analysis of myosin IIIA containing the kinase domain (MIII). The steady-state kinetic properties of MIII ΔK indicate that it has a 2-fold higher maximum actin-activated ATPase rate (kcat = 1.5 ± 0.1 s-1) and a 5-fold tighter actin affinity (KATPase = 6.0 ± 1.4 μM, and KActin = 1.4 ± 0.4 μM) compared to MIII. The rate of ATP binding to the motor domain is enhanced in MIII ΔK (K1k+2 ≈ 0.10 ± 0.01 μM -1·s-1) to a level similar to the rate of binding to MIII in the presence of actin. The rate of ATP hydrolysis in the absence of actin is slow and may be rate limiting. Actin-activated phosphate release is identical with and without the kinase domain. The transition between actomyosin.ADP states, which is rate limiting in MIII, is enhanced in MIII ΔK. MIII ΔK accumulates more efficiently at the tips of filopodia in HeLa cells. Our results suggest a model in which the activity and concentration of myosin IIIA localized to the tips of actin bundles mediates the morphology of the tips in sensory cells.

Original languageEnglish (US)
Pages (from-to)2485-2496
Number of pages12
JournalBiochemistry
Volume47
Issue number8
DOIs
StatePublished - Feb 26 2008

Fingerprint

Myosins
Phosphotransferases
Actins
Kinetics
Inner Auditory Hair Cells
Adenosine Triphosphate
Stereocilia
Actomyosin
Pseudopodia
Inner Ear
HeLa Cells
Adenosine Diphosphate
Hydrolysis
Phosphates
Cells
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Dosé, A. C., Ananthanarayanan, S., Moore, J. E., Corsa, A. C., Burnside, B., & Yengo, C. (2008). The kinase domain alters the kinetic properties of the myosin IIIA motor. Biochemistry, 47(8), 2485-2496. https://doi.org/10.1021/bi7021574
Dosé, Andréa C. ; Ananthanarayanan, Shobana ; Moore, Judy E. ; Corsa, Amoreena C. ; Burnside, Beth ; Yengo, Christopher. / The kinase domain alters the kinetic properties of the myosin IIIA motor. In: Biochemistry. 2008 ; Vol. 47, No. 8. pp. 2485-2496.
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Dosé, AC, Ananthanarayanan, S, Moore, JE, Corsa, AC, Burnside, B & Yengo, C 2008, 'The kinase domain alters the kinetic properties of the myosin IIIA motor', Biochemistry, vol. 47, no. 8, pp. 2485-2496. https://doi.org/10.1021/bi7021574

The kinase domain alters the kinetic properties of the myosin IIIA motor. / Dosé, Andréa C.; Ananthanarayanan, Shobana; Moore, Judy E.; Corsa, Amoreena C.; Burnside, Beth; Yengo, Christopher.

In: Biochemistry, Vol. 47, No. 8, 26.02.2008, p. 2485-2496.

Research output: Contribution to journalArticle

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AU - Dosé, Andréa C.

AU - Ananthanarayanan, Shobana

AU - Moore, Judy E.

AU - Corsa, Amoreena C.

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AU - Yengo, Christopher

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Y1 - 2008/2/26

N2 - Myosin IIIA is unique among myosin proteins in that it contains an N-terminal kinase domain capable of autophosphorylating sites on the motor domain. A construct of myosin IIIA lacking the kinase domain localizes more efficiently to the stereocilia tips and alters the morphology of the tips in inner ear hair cells. Therefore, we performed a kinetic analysis of myosin IIIA without the kinase domain (MIII ΔK) and compared these results with our reported analysis of myosin IIIA containing the kinase domain (MIII). The steady-state kinetic properties of MIII ΔK indicate that it has a 2-fold higher maximum actin-activated ATPase rate (kcat = 1.5 ± 0.1 s-1) and a 5-fold tighter actin affinity (KATPase = 6.0 ± 1.4 μM, and KActin = 1.4 ± 0.4 μM) compared to MIII. The rate of ATP binding to the motor domain is enhanced in MIII ΔK (K1k+2 ≈ 0.10 ± 0.01 μM -1·s-1) to a level similar to the rate of binding to MIII in the presence of actin. The rate of ATP hydrolysis in the absence of actin is slow and may be rate limiting. Actin-activated phosphate release is identical with and without the kinase domain. The transition between actomyosin.ADP states, which is rate limiting in MIII, is enhanced in MIII ΔK. MIII ΔK accumulates more efficiently at the tips of filopodia in HeLa cells. Our results suggest a model in which the activity and concentration of myosin IIIA localized to the tips of actin bundles mediates the morphology of the tips in sensory cells.

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Dosé AC, Ananthanarayanan S, Moore JE, Corsa AC, Burnside B, Yengo C. The kinase domain alters the kinetic properties of the myosin IIIA motor. Biochemistry. 2008 Feb 26;47(8):2485-2496. https://doi.org/10.1021/bi7021574