Integrative biology of native cell extracts : a new era for structural characterization of life processes

Advances in electron microscopy have provided unprecedented access to the structural characterization of large, flexible and heterogeneous complexes. Until recently, cryo-electron microscopy (cryo-EM) has been applied to understand molecular organization in either highly purified, isolated biomolecules or in situ. An emerging field is developing, bridging the gap between the two approaches, and focuses on studying molecular organization in native cell extracts. This field has demonstrated its potential by resolving the structure of fungal fatty acid synthase (FAS) at 4.7 Å [Fourier shell correlation (FSC) = 0.143]; FAS was not only less than 50% enriched, but also retained higher-order binders, previously unknown. Although controversial in the sense that the lysis step might introduce artifacts, cell extracts preserve aspects of cellular function. In addition, cell extracts are accessible, besides cryo-EM, to modern proteomic methods, chemical cross-linking, network biology and biophysical modeling. We expect that automation in imaging cell extracts, along with the integration of molecular/cell biology approaches, will provide remarkable achievements in the study of closer-to-life biomolecular states of pronounced biotechnological and medical importance. Such steps will, eventually, bring us a step closer to the biophysical description of cellular processes in an integrative, holistic approach.

Medienart:

E-Artikel

Erscheinungsjahr:

2019

Erschienen:

2019

Enthalten in:

Zur Gesamtaufnahme - volume:400

Enthalten in:

Biological chemistry - 400(2019), 7 vom: 26. Juni, Seite 831-846

Sprache:

Englisch

Beteiligte Personen:

Kyrilis, Fotis L [VerfasserIn]
Meister, Annette [VerfasserIn]
Kastritis, Panagiotis L [VerfasserIn]

Links:

Volltext

Themen:

Biomolecular interactions
Cell Extracts
Cell-free systems
Cryo-EM
Flexible docking
Image processing
Journal Article
Proteins
Research Support, Non-U.S. Gov't
Review
XL-MS

Anmerkungen:

Date Completed 17.02.2020

Date Revised 12.02.2021

published: Print

Citation Status MEDLINE

doi:

10.1515/hsz-2018-0445

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM297098993