Increasing evidence of mechanical force as a functional regulator in smooth muscle myosin light chain kinase

Fabian Baumann; Magnus Sebastian Bauer; Martin Rees; Alexander Alexandrovich; Mathias Gautel; Diana Angela Pippig; Hermann Eduard Gaub

doi:10.7554/eLife.26473

Increasing evidence of mechanical force as a functional regulator in smooth muscle myosin light chain kinase

Fabian Baumann, Magnus Sebastian Bauer, Martin Rees, Alexander Alexandrovich, Mathias Gautel, Diana Angela Pippig, Hermann Eduard Gaub^*

^*Corresponding author for this work

Pettenkofer School of Public Health

Research output: Contribution to journal › Article › peer-review

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Abstract

Mechanosensitive proteins are key players in cytoskeletal remodeling, muscle contraction, cell migration and differentiation processes. Smooth muscle myosin light chain kinase (smMLCK) is a member of a diverse group of serine/threonine kinases that feature cytoskeletal association. Its catalytic activity is triggered by a conformational change upon Ca²⁺/calmodulin (Ca²⁺/CaM) binding. Due to its significant homology with the force-activated titin kinase, smMLCK is suspected to be also regulatable by mechanical stress. In this study, a CaM-independent activation mechanism for smMLCK by mechanical release of the inhibitory elements is investigated via high throughput AFM single-molecule force spectroscopy. The characteristic pattern of transitions between different smMLCK states and their variations in the presence of different substrates and ligands are presented. Interaction between kinase domain and regulatory light chain (RLC) substrate is identified in the absence of CaM, indicating restored substrate-binding capability due to mechanically induced removal of the auto-inhibitory regulatory region.

Original language	English
Article number	e26473
Journal	eLife
Volume	6
DOIs	https://doi.org/10.7554/eLife.26473
Publication status	Published - 11 Jul 2017

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Increasing evidence of mechanical_BAUMANN_Publishedonline11July2017_GOLD VoR (CC BY)Final published version, 1.13 MBLicence: CC BY

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abstract = "Mechanosensitive proteins are key players in cytoskeletal remodeling, muscle contraction, cell migration and differentiation processes. Smooth muscle myosin light chain kinase (smMLCK) is a member of a diverse group of serine/threonine kinases that feature cytoskeletal association. Its catalytic activity is triggered by a conformational change upon Ca2+/calmodulin (Ca2+/CaM) binding. Due to its significant homology with the force-activated titin kinase, smMLCK is suspected to be also regulatable by mechanical stress. In this study, a CaM-independent activation mechanism for smMLCK by mechanical release of the inhibitory elements is investigated via high throughput AFM single-molecule force spectroscopy. The characteristic pattern of transitions between different smMLCK states and their variations in the presence of different substrates and ligands are presented. Interaction between kinase domain and regulatory light chain (RLC) substrate is identified in the absence of CaM, indicating restored substrate-binding capability due to mechanically induced removal of the auto-inhibitory regulatory region.",

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AU - Baumann, Fabian

AU - Bauer, Magnus Sebastian

AU - Rees, Martin

AU - Alexandrovich, Alexander

AU - Gautel, Mathias

AU - Pippig, Diana Angela

AU - Gaub, Hermann Eduard

PY - 2017/7/11

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AB - Mechanosensitive proteins are key players in cytoskeletal remodeling, muscle contraction, cell migration and differentiation processes. Smooth muscle myosin light chain kinase (smMLCK) is a member of a diverse group of serine/threonine kinases that feature cytoskeletal association. Its catalytic activity is triggered by a conformational change upon Ca2+/calmodulin (Ca2+/CaM) binding. Due to its significant homology with the force-activated titin kinase, smMLCK is suspected to be also regulatable by mechanical stress. In this study, a CaM-independent activation mechanism for smMLCK by mechanical release of the inhibitory elements is investigated via high throughput AFM single-molecule force spectroscopy. The characteristic pattern of transitions between different smMLCK states and their variations in the presence of different substrates and ligands are presented. Interaction between kinase domain and regulatory light chain (RLC) substrate is identified in the absence of CaM, indicating restored substrate-binding capability due to mechanically induced removal of the auto-inhibitory regulatory region.

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Increasing evidence of mechanical force as a functional regulator in smooth muscle myosin light chain kinase

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