Intermediate filaments (IFs), along with microtubules, microfilaments, and associated cross-bridging proteins, constitute the cytoskeleton of metazoan cells. While crystallographic data on the dimer representing the elementary IF "building block" have recently become available, little structural detail is known about both the mature IF architecture and its assembly pathway. Here, we have applied solution small-angle x-ray scattering to investigate the in vitro assembly of a 53-kDa human IF protein vimentin at pH 8.4 by systematically varying the ionic strength conditions, and complemented these experiments by electron microscopy and analytical ultracentrifugation. While a vimentin solution in 5 mM Tris.HCl (pH 8.4) contains predominantly tetramers, addition of 20 mM NaCl induces further lateral assembly evidenced by the shift of the sedimentation coefficient and yields a distinct octameric intermediate. Four octamers eventually associate into unit-length filaments (ULFs) that anneal longitudinally. Based on the small-angle x-ray scattering experiments supplemented by crystallographic data and additional structural constraints, 3D molecular models of the vimentin tetramer, octamer, and ULF were constructed. Within each of the three oligomers, the adjacent dimers are aligned exclusively in an approximately half-staggered antiparallel A(11) mode with a distance of 3.2-3.4 nm between their axes. The ULF appears to be a dynamic and a relatively loosely packed structure with a roughly even mass distribution over its cross-section.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1637561PMC
http://dx.doi.org/10.1073/pnas.0603629103DOI Listing

Publication Analysis

Top Keywords

small-angle x-ray
12
x-ray scattering
12
architecture assembly
8
crystallographic data
8
assembly
5
monitoring intermediate
4
intermediate filament
4
filament assembly
4
assembly small-angle
4
scattering reveals
4

Similar Publications

Structural Transformation and Degradation of Cu Oxide Nanocatalysts during Electrochemical CO Reduction.

J Am Chem Soc

January 2025

Liquid Sunlight Alliance, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, California 94720, United States.

The electrochemical CO reduction reaction (CORR) holds enormous potential as a carbon-neutral route to the sustainable production of fuels and platform chemicals. The durability for long-term operation is currently inadequate for commercialization, however, and the underlying deactivation process remains elusive. A fundamental understanding of the degradation mechanism of electrocatalysts, which can dictate the overall device performance, is needed.

View Article and Find Full Text PDF

The structural organisation of pentraxin-3 and its interactions with heavy chains of inter-α-inhibitor regulate crosslinking of the hyaluronan matrix.

Matrix Biol

January 2025

Manchester Cell-Matrix Centre, Division of Cell-Matrix Biology and Regenerative Medicine, School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester Academic Health Science Centre, Manchester M13 9PT, UK; Lydia Becker Institute of Immunology and Inflammation, University of Manchester, Manchester, M13 9PL, United Kingdom. Electronic address:

Pentraxin-3 (PTX3) is an octameric protein, comprised of eight identical protomers, that has diverse functions in reproductive biology, innate immunity and cancer. PTX3 interacts with the large polysaccharide hyaluronan (HA) to which heavy chains (HCs) of the inter-α-inhibitor (IαI) family of proteoglycans are covalently attached, playing a key role in the (non-covalent) crosslinking of HC•HA complexes. These interactions stabilise the cumulus matrix, essential for ovulation and fertilisation in mammals, and are also implicated in the formation of pathogenic matrices in the context of viral lung infections.

View Article and Find Full Text PDF

Electrochemical Small-Angle X-ray Scattering for Potential-Dependent Structural Analysis of Redox Enzymes.

Langmuir

January 2025

Department of Pure and Applied Chemistry, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.

Various methods exist for exploring different aspects of these mechanisms. However, techniques for investigating structural differences between the reduced and oxidized forms of an enzyme are limited. Here, we propose electrochemical small-angle X-ray scattering (EC-SAXS) as a novel method for potential-dependent structural analysis of redox enzymes and redox-active proteins.

View Article and Find Full Text PDF

All-solid-state (ASS) batteries are a promising solution to achieve carbon neutrality. ASS lithium-sulfur (Li-S) batteries stand out due to their improved safety, achieved by replacing organic solvents, which are prone to leakage and fire, with solid electrolytes. In addition, these batteries offer the benefits of higher capacity and the absence of rare metals.

View Article and Find Full Text PDF

Mitochondrial metabolism requires the chaperoned import of disulfide-stabilized proteins via CHCHD4/MIA40 and its enigmatic interaction with oxidoreductase Apoptosis-inducing factor (AIF). By crystallizing human CHCHD4's AIF-interaction domain with an activated AIF dimer, we uncover how NADH allosterically configures AIF to anchor CHCHD4's β-hairpin and histidine-helix motifs to the inner mitochondrial membrane. The structure further reveals a similarity between the AIF-interaction domain and recognition sequences of CHCHD4 substrates.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!