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Cytoskeletal filaments are polymeric forms of smaller protein subunits. However, individual cytoskeletal filaments may easily disassemble or associate with other similar filaments to form rigid structures. Microfilaments, made of actin monomers, rely on actin-binding proteins to form bundles and create networks of individual actin filaments. Microtubules rely on microtubule-associated proteins (MAPs) to form sturdy cylindrical structures. However, the proteins involved in forming complex...
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Synthetic biology is an interdisciplinary science that involves using principles from disciplines such as engineering, molecular biology, cell biology, and systems biology. It involves remodeling existing organisms from nature or constructing completely new synthetic organisms for applications such as protein or enzyme production, bioremediation, value-added macromolecule production, and the addition of desirable traits to crops, to name a few.
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Anchoring junctions are multiprotein complexes that help cells connect to other cells and the extracellular matrix. Anchoring junctions are present on the lateral and basal surfaces of cells, providing strong and flexible connections. Focal adhesions are often formed due to cell interactions with the ECM substrata, which initiate signal transduction via kinase cascades and other mechanisms. Together, they provide stability and tissue integrity. There are three types of anchoring junctions:...
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The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...
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Structural biology in cellulo: Minding the gap between conceptualization and realization.

Fotis L Kyrilis1, Jason K K Low2, Joel P Mackay2

  • 1Institute of Chemical Biology, National Hellenic Research Foundation, Athens, Greece. Electronic address: https://twitter.com/Fotansky_16.

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Advancing cellular structural biology requires integrating multiple imaging and biochemical techniques. Future efforts must expand beyond proteins to encompass all essential macromolecules for a complete cellular picture.

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Area of Science:

  • Cellular Biology
  • Structural Biology
  • Biochemistry

Background:

  • Current structural biology methods often analyze purified components, not intact cells.
  • This limitation hinders a comprehensive understanding of cellular processes and organization.

Purpose of the Study:

  • To review current and emerging technologies for in-cell structural biology.
  • To highlight the need for integrating diverse approaches for a holistic cellular view.

Main Methods:

  • Electron cryotomography (cryo-ET) for in cellulo snapshots.
  • Single-particle electron microscopy (SP-EM) for purified and complex mixtures.
  • Covalent crosslinking with mass spectrometry (XL-MS).
  • Artificial intelligence (AI) algorithms for data analysis.

Main Results:

  • Cryo-ET provides high-resolution snapshots of cellular structures in their native state.
  • SP-EM and XL-MS offer complementary structural insights.
  • AI enhances the interpretation and integration of complex structural datasets.

Conclusions:

  • Integrating cryo-ET, SP-EM, XL-MS, and AI is crucial for advancing in-cell structural biology.
  • Expanding focus beyond proteins to other macromolecules is essential for a complete understanding of cellular life.