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佛罗里达木匠蚁 Camponotus floridanus 的下颌肌肌钙蛋白:扩展我们对无脊椎动物 Ca 调节的认识。

Mandibular muscle troponin of the Florida carpenter ant Camponotus floridanus: extending our insights into invertebrate Ca regulation.

机构信息

Department of Biological Science, Florida State University, Tallahassee, FL, 32306, USA.

Department of Biomedical Sciences, Florida State University College of Medicine, Tallahassee, FL, 32306, USA.

出版信息

J Muscle Res Cell Motil. 2021 Jun;42(2):399-417. doi: 10.1007/s10974-021-09606-w. Epub 2021 Jul 13.

Abstract

Ants use their mandibles for a variety of functions and behaviors. We investigated mandibular muscle structure and function from major workers of the Florida carpenter ant Camponotus floridanus: force-pCa relation and velocity of unloaded shortening of single, permeabilized fibres, primary sequences of troponin subunits (TnC, TnI and TnT) from a mandibular muscle cDNA library, and muscle fibre ultrastructure. From the mechanical measurements, we found Ca-sensitivity of isometric force was markedly shifted rightward compared with vertebrate striated muscle. From the troponin sequence results, we identified features that could explain the rightward shift of Ca-activation: the N-helix of TnC is effectively absent and three of the four EF-hands of TnC (sites I, II and III) do not adhere to canonical sequence rules for divalent cation binding; two alternatively spliced isoforms of TnI were identified with the alternatively spliced exon occurring in the region of the IT-arm α-helical coiled-coil, and the N-terminal extension of TnI may be involved in modulation of regulation, as in mammalian cardiac muscle; and TnT has a Glu-rich C-terminus. In addition, a structural homology model was built of C. floridanus troponin on the thin filament. From analysis of electron micrographs, we found thick filaments are almost as long as the 6.8 μm sarcomeres, have diameter of ~ 16 nm, and typical center-to-center spacing of ~ 46 nm. These results have implications for the mechanisms by which mandibular muscle fibres perform such a variety of functions, and how the structure of the troponin complex aids in these tasks.

摘要

蚂蚁用它们的下颚执行各种功能和行为。我们研究了佛罗里达木匠蚁 Camponotus floridanus 的工蚁下颚的肌肉结构和功能:单根渗透化纤维的力-pCa 关系和无负荷缩短速度、下颚肌肉 cDNA 文库中的肌钙蛋白亚基(TnC、TnI 和 TnT)的原始序列以及肌肉纤维超微结构。从力学测量结果中,我们发现与脊椎动物横纹肌相比,等长力的 Ca 敏感性明显右移。从肌钙蛋白序列结果中,我们确定了可以解释 Ca 激活右移的特征:TnC 的 N 螺旋有效缺失,并且 TnC 的四个 EF 手(位点 I、II 和 III)中的三个不遵守二价阳离子结合的经典序列规则;鉴定出两种 TnI 的选择性剪接同工型,其中选择性剪接外显子发生在 IT 臂α螺旋卷曲螺旋区,并且 TnI 的 N 端延伸可能参与调节,就像在哺乳动物心肌中一样;并且 TnT 具有富含 Glu 的 C 末端。此外,还在细丝上构建了 C. floridanus 肌钙蛋白的结构同源模型。从电子显微镜照片分析中,我们发现粗丝几乎与 6.8 μm 的肌节一样长,直径约为 16nm,并且具有典型的中心到中心间隔约为 46nm。这些结果对下颚肌肉纤维执行各种功能的机制以及肌钙蛋白复合物的结构如何帮助完成这些任务具有重要意义。

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