• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

越来越多的证据表明机械力是平滑肌肌球蛋白轻链激酶的一种功能调节因子。

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

作者信息

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

机构信息

Chair for Applied Physics and Center for Nanoscience, Ludwig-Maximilians-Universität München, Munich, Germany.

Center for Integrated Protein Science Munich, Ludwig-Maximilians-Universität München, Munich, Germany.

出版信息

Elife. 2017 Jul 11;6:e26473. doi: 10.7554/eLife.26473.

DOI:10.7554/eLife.26473
PMID:28696205
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5505704/
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 Ca/calmodulin (Ca/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.

摘要

机械敏感蛋白是细胞骨架重塑、肌肉收缩、细胞迁移和分化过程中的关键参与者。平滑肌肌球蛋白轻链激酶（smMLCK）是一组具有细胞骨架关联特性的丝氨酸/苏氨酸激酶中的一员。其催化活性在钙/钙调蛋白（Ca/CaM）结合后通过构象变化触发。由于其与力激活的肌联蛋白激酶具有显著同源性，推测smMLCK也可由机械应力调节。在本研究中，通过高通量原子力显微镜单分子力谱研究了通过抑制元件的机械释放实现的smMLCK的钙调蛋白非依赖性激活机制。呈现了不同smMLCK状态之间转变的特征模式及其在不同底物和配体存在下的变化。在没有钙调蛋白的情况下鉴定了激酶结构域与调节轻链（RLC）底物之间的相互作用，表明由于机械诱导去除自抑制调节区域而恢复了底物结合能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/4f351d518b4c/elife-26473-fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/ba1bb0fc9d68/elife-26473-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/dd58473857b9/elife-26473-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/427fe69b7c1c/elife-26473-fig2-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/19bbf078eda6/elife-26473-fig2-figsupp2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/0c165ae75e96/elife-26473-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/dd03d68adeb5/elife-26473-fig3-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/dbb07947c37b/elife-26473-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/bbbde59bbaa4/elife-26473-fig4-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/4f351d518b4c/elife-26473-fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/ba1bb0fc9d68/elife-26473-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/dd58473857b9/elife-26473-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/427fe69b7c1c/elife-26473-fig2-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/19bbf078eda6/elife-26473-fig2-figsupp2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/0c165ae75e96/elife-26473-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/dd03d68adeb5/elife-26473-fig3-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/dbb07947c37b/elife-26473-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/bbbde59bbaa4/elife-26473-fig4-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73ac/5505704/4f351d518b4c/elife-26473-fig5.jpg

相似文献

1
Increasing evidence of mechanical force as a functional regulator in smooth muscle myosin light chain kinase.越来越多的证据表明机械力是平滑肌肌球蛋白轻链激酶的一种功能调节因子。
Elife. 2017 Jul 11;6:e26473. doi: 10.7554/eLife.26473.
2
Myosin light chain kinase steady-state kinetics: comparison of smooth muscle myosin II and nonmuscle myosin IIB as substrates.肌球蛋白轻链激酶稳态动力学：平滑肌肌球蛋白II和非肌肉肌球蛋白IIB作为底物的比较
Cell Biochem Funct. 2016 Oct;34(7):469-474. doi: 10.1002/cbf.3209. Epub 2016 Aug 16.
3
Calmodulin kinase II chimeras used to investigate the structural requirements for smooth muscle myosin light chain kinase autoinhibition and calmodulin-dependent activation.用于研究平滑肌肌球蛋白轻链激酶自身抑制和钙调蛋白依赖性激活的结构要求的钙调蛋白激酶II嵌合体。
Biochemistry. 1999 Nov 16;38(46):15061-9. doi: 10.1021/bi990883a.
4
Myosin light chain kinase activation and calcium sensitization in smooth muscle in vivo.体内平滑肌中的肌球蛋白轻链激酶激活与钙敏化
Am J Physiol Cell Physiol. 2008 Aug;295(2):C358-64. doi: 10.1152/ajpcell.90645.2007. Epub 2008 Jun 4.
5
Real-time evaluation of myosin light chain kinase activation in smooth muscle tissues from a transgenic calmodulin-biosensor mouse.对来自转基因钙调蛋白生物传感器小鼠的平滑肌组织中肌球蛋白轻链激酶激活的实时评估。
Proc Natl Acad Sci U S A. 2004 Apr 20;101(16):6279-84. doi: 10.1073/pnas.0308742101. Epub 2004 Apr 7.
6
In vivo roles for myosin phosphatase targeting subunit-1 phosphorylation sites T694 and T852 in bladder smooth muscle contraction.肌球蛋白磷酸酶靶向亚基-1磷酸化位点T694和T852在膀胱平滑肌收缩中的体内作用。
J Physiol. 2015 Feb 1;593(3):681-700. doi: 10.1113/jphysiol.2014.283853. Epub 2014 Dec 23.
7
Kinetics of myosin light chain kinase activation of smooth muscle myosin in an in vitro model system.在体外模型系统中肌球蛋白轻链激酶对平滑肌肌球蛋白的激活动力学。
Biochemistry. 2013 Nov 26;52(47):8489-500. doi: 10.1021/bi401001x. Epub 2013 Nov 11.
8
Physiological vs. pharmacological signalling to myosin phosphorylation in airway smooth muscle.气道平滑肌肌球蛋白磷酸化的生理与药理学信号转导。
J Physiol. 2017 Oct 1;595(19):6231-6247. doi: 10.1113/JP274715. Epub 2017 Aug 24.
9
Physiological signalling to myosin phosphatase targeting subunit-1 phosphorylation in ileal smooth muscle.回肠平滑肌中肌球蛋白磷酸酶靶向亚基-1磷酸化的生理信号传导
J Physiol. 2016 Jun 15;594(12):3209-25. doi: 10.1113/JP271703. Epub 2016 Mar 4.
10
Enhanced skeletal muscle contraction with myosin light chain phosphorylation by a calmodulin-sensing kinase.钙调蛋白传感激酶通过肌球蛋白轻链磷酸化增强骨骼肌收缩。
J Biol Chem. 2007 Jul 13;282(28):20447-54. doi: 10.1074/jbc.M702927200. Epub 2007 May 15.

引用本文的文献

1
Tension can directly suppress Aurora B kinase-triggered release of kinetochore-microtubule attachments.紧张会直接抑制 Aurora B 激酶触发的动粒微管附着的释放。
Nat Commun. 2022 Apr 20;13(1):2152. doi: 10.1038/s41467-022-29542-8.
2
A tethered ligand assay to probe SARS-CoV-2:ACE2 interactions.一种用于探测 SARS-CoV-2:ACE2 相互作用的连接配体测定法。
Proc Natl Acad Sci U S A. 2022 Apr 5;119(14):e2114397119. doi: 10.1073/pnas.2114397119. Epub 2022 Mar 21.
3
Hypoxic modulation of fetal vascular MLCK abundance, localization, and function.

本文引用的文献

1
Cardiac myosin light chain is phosphorylated by Ca2+/calmodulin-dependent and -independent kinase activities.心肌肌球蛋白轻链通过钙/钙调蛋白依赖性和非依赖性激酶活性被磷酸化。
Proc Natl Acad Sci U S A. 2016 Jul 5;113(27):E3824-33. doi: 10.1073/pnas.1600633113. Epub 2016 Jun 20.
2
Monovalent Strep-Tactin for strong and site-specific tethering in nanospectroscopy.单价链霉亲合素在纳米光谱学中用于强和特异性固定。
Nat Nanotechnol. 2016 Jan;11(1):89-94. doi: 10.1038/nnano.2015.231. Epub 2015 Oct 12.
3
Diffusion of myosin light chain kinase on actin: A mechanism to enhance myosin phosphorylation rates in smooth muscle.
低氧对胎儿血管 MLCK 丰度、定位和功能的调节作用。
Am J Physiol Regul Integr Comp Physiol. 2021 Jan 1;320(1):R1-R18. doi: 10.1152/ajpregu.00212.2020. Epub 2020 Oct 28.
4
Effects of Ligand Binding on the Energy Landscape of Acyl-CoA-Binding Protein.配体结合对酰基辅酶A结合蛋白能量景观的影响。
Biophys J. 2020 Nov 3;119(9):1821-1832. doi: 10.1016/j.bpj.2020.09.016. Epub 2020 Sep 24.
5
Next Generation Methods for Single-Molecule Force Spectroscopy on Polyproteins and Receptor-Ligand Complexes.多聚蛋白和受体-配体复合物单分子力谱的下一代方法
Front Mol Biosci. 2020 May 19;7:85. doi: 10.3389/fmolb.2020.00085. eCollection 2020.
6
Structural and mechanistic insights into mechanoactivation of focal adhesion kinase.机械激活粘着斑激酶的结构和机制见解。
Proc Natl Acad Sci U S A. 2019 Apr 2;116(14):6766-6774. doi: 10.1073/pnas.1820567116. Epub 2019 Mar 15.
7
The myosin light-chain kinase MLCK-1 relocalizes during Caenorhabditis elegans ovulation to promote actomyosin bundle assembly and drive contraction.肌球蛋白轻链激酶 MLCK-1 在 Caenorhabditis elegans 排卵期间重新定位，以促进肌动球蛋白束的组装并推动收缩。
Mol Biol Cell. 2018 Aug 8;29(16):1975-1991. doi: 10.1091/mbc.E18-01-0056. Epub 2018 Apr 5.
8
Combinatorial Contact Cues Specify Cell Division Orientation by Directing Cortical Myosin Flows.组合接触线索通过指导皮质肌球蛋白流来指定细胞分裂方向。
Dev Cell. 2018 Aug 6;46(3):257-270.e5. doi: 10.1016/j.devcel.2018.06.020. Epub 2018 Jul 19.
肌球蛋白轻链激酶在肌动蛋白上的扩散：一种提高平滑肌中肌球蛋白磷酸化速率的机制。
J Gen Physiol. 2015 Oct;146(4):267-80. doi: 10.1085/jgp.201511483.
4
Mechanobiology.机械生物学
Integr Biol (Camb). 2015 Oct;7(10):1091-2. doi: 10.1039/c5ib90040a. Epub 2015 Sep 25.
5
Myosin light chain kinase (MLCK) regulates cell migration in a myosin regulatory light chain phosphorylation-independent mechanism.肌球蛋白轻链激酶（MLCK）通过一种不依赖于肌球蛋白调节性轻链磷酸化的机制来调控细胞迁移。
J Biol Chem. 2014 Oct 10;289(41):28478-88. doi: 10.1074/jbc.M114.567446. Epub 2014 Aug 13.
6
Investigating receptor-ligand systems of the cellulosome with AFM-based single-molecule force spectroscopy.利用基于原子力显微镜的单分子力谱研究纤维小体的受体-配体系统。
J Vis Exp. 2013 Dec 20(82):e50950. doi: 10.3791/50950.
7
Nonspecific interactions in AFM force spectroscopy measurements.原子力显微镜力谱测量中的非特异性相互作用。
J Mol Recognit. 2012 Jan;25(1):53-6. doi: 10.1002/jmr.2152.
8
Biochemistry of smooth muscle myosin light chain kinase.平滑肌肌球蛋白轻链激酶的生物化学
Arch Biochem Biophys. 2011 Jun 15;510(2):135-46. doi: 10.1016/j.abb.2011.04.018. Epub 2011 May 3.
9
Cytoskeletal protein kinases: titin and its relations in mechanosensing.细胞骨架蛋白激酶：titin 及其在机械感知中的关系。
Pflugers Arch. 2011 Jul;462(1):119-34. doi: 10.1007/s00424-011-0946-1. Epub 2011 Mar 18.
10
Thiol-based, site-specific and covalent immobilization of biomolecules for single-molecule experiments.基于巯基的、针对特定位置的、生物分子的共价固定化用于单分子实验。
Nat Protoc. 2010 Jun;5(6):975-85. doi: 10.1038/nprot.2010.49.