skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

Explore Research Products in the PAR
It may take a few hours for recently added research products to appear in PAR search results.


  • NSF Public Access
  • Search Results
  • Enzymatic noncovalent synthesis of peptide assemblies generates multimolecular crowding in cells for biomedical applications
Title: Enzymatic noncovalent synthesis of peptide assemblies generates multimolecular crowding in cells for biomedical applications
Enzymatic noncovalent synthesis enables the spatiotemporal control of multimolecular crowding in cells, thus offering a unique opportunity for modulating cellular functions. This article introduces some representative enzymes and molecular building blocks for generating peptide assemblies as multimolecular crowding in cells, highlights the relevant biomedical applications, such as anticancer therapy, molecular imaging, trafficking proteins, genetic engineering, artificial intracellular filaments, cell morphogenesis, and antibacterial, and briefly discusses the promises of ENS as a multistep molecular process in biology and medicine.  more » « less
Award ID(s):
2011846
PAR ID:
10506786
Author(s) / Creator(s):
; ; ;
Publisher / Repository:
Royal Society of Chemistry
Date Published:
Journal Name:
Chemical Communications
Volume:
57
Issue:
96
ISSN:
1359-7345
Page Range / eLocation ID:
12870 to 12879
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. ; ; ; ; (, Biomolecules)
    Actin polymerization drives cell movement and provides cells with structural integrity. Intracellular environments contain high concentrations of solutes, including organic compounds, macromolecules, and proteins. Macromolecular crowding has been shown to affect actin filament stability and bulk polymerization kinetics. However, the molecular mechanisms behind how crowding influences individual actin filament assembly are not well understood. In this study, we investigated how crowding modulates filament assembly kinetics using total internal reflection fluorescence (TIRF) microscopy imaging and pyrene fluorescence assays. The elongation rates of individual actin filaments analyzed from TIRF imaging depended on the type of crowding agent (polyethylene glycol, bovine serum albumin, and sucrose) as well as their concentrations. Further, we utilized all-atom molecular dynamics (MD) simulations to evaluate the effects of crowding molecules on the diffusion of actin monomers during filament assembly. Taken together, our data suggest that solution crowding can regulate actin assembly kinetics at the molecular level. 
    more » « less
  2. ; ; ; (, Proceedings of the National Academy of Sciences)
    Cells mediate interactions with the extracellular environment through a crowded assembly of transmembrane proteins, glycoproteins and glycolipids on their plasma membrane. The extent to which surface crowding modulates the biophysical interactions of ligands, receptors, and other macromolecules is poorly understood due to the lack of methods to quantify surface crowding on native cell membranes. In this work, we demonstrate that physical crowding on reconstituted membranes and live cell surfaces attenuates the effective binding affinity of macromolecules such as IgG antibodies in a surface crowding-dependent manner. We combine experiment and simulation to design a crowding sensor based on this principle that provides a quantitative readout of cell surface crowding. Our measurements reveal that surface crowding decreases IgG antibody binding by 2 to 20 fold in live cells compared to a bare membrane surface. Our sensors show that sialic acid, a negatively charged monosaccharide, contributes disproportionately to red blood cell surface crowding via electrostatic repulsion, despite occupying only ~1% of the total cell membrane by mass. We also observe significant differences in surface crowding for different cell types and find that expression of single oncogenes can both increase and decrease crowding, suggesting that surface crowding may be an indicator of both cell type and state. Our high-throughput, single-cell measurement of cell surface crowding may be combined with functional assays to enable further biophysical dissection of the cell surfaceome. 
    more » « less
  3. ; ; ; ; ; ; (, Biomaterials Science)
    Collagen I is the primary extracellular matrix component of most solid tumors and influences metastatic progression. Collagen matrix engineering techniques are useful for understanding how this complex biomaterial regulates cancer cell behavior and for improving in vitro cancer models. Here, we establish an approach to tune collagen fibril architecture using PEG as an inert molecular crowding agent during gelation and cell embedding. We find that crowding produces matrices with tighter fibril networks that are less susceptible to proteinase mediated degradation, but does not significantly alter matrix stiffness. The resulting matrices have the effect of preventing cell spreading, confining cells, and reducing cell contractility. Matrix degradability and fibril length are identified as strong predictors of cell confinement. Further, the degree of confinement predicts whether breast cancer cells will ultimately undergo individual or collective behaviors. Highly confined breast cancer cells undergo morphogenesis to form either invasive networks reminiscent of aggressive tumors or gland and lobule structures reminiscent of normal breast epithelia. This morphological transition is accompanied by expression of cell–cell adhesion genes, including PECAM1 and ICAM1. Our study suggests that cell confinement, mediated by matrix architecture, is a design feature that tunes the transcriptional and morphogenic state of breast cancer cells. 
    more » « less
  4. ; ; ; ; ; ; (, Journal of The Electrochemical Society)
    Aqueous Li-ion batteries (ALIBs) are an important class of battery chemistries owing to the intrinsic non-flammability of aqueous electrolytes. However, water is detrimental to most cathode materials and could result in rapid cell failure. Identifying the degradation mechanisms and evaluating the pros and cons of different cathode materials are crucial to guide the materials selection and maximize their electrochemical performance in ALIBs. In this study, we investigate the stability of LiFePO4(LFP), LiMn2O4(LMO) and LiNi0.8Mn0.1Co0.1O2(NMC) cathodes, without protective coating, in three different aqueous electrolytes, i.e., salt-in-water, water-in-salt, and molecular crowding electrolytes. The latter two are the widely reported “water-deficient electrolytes.” LFP cycled in the molecular crowding electrolyte exhibits the best cycle life in both symmetric and full cells owing to the stable crystal structure. Mn dissolution and surface reduction accelerate the capacity decay of LMO in water-rich electrolyte. On the other hand, the bulk structural collapse leads to the degradation of NMC cathodes. LMO demonstrates better full-cell performance than NMC in water-deficient aqueous electrolytes. LFP is shown to be more promising than LMO and NMC for long-cycle-life ALIB full cells, especially in the molecular crowding electrolyte. However, none of the aqueous electrolytes studied here provide enough battery performance that can compete with conventional non-aqueous electrolytes. This work reveals the degradation mechanisms of olivine, spinel, and layered cathodes in different aqueous electrolytes and yields insights into improving electrode materials and electrolytes for ALIBs. 
    more » « less
  5. ; ; (, The Journal of Chemical Physics)
    The diffusion of proteins is significantly affected by macromolecular crowding. Molecular simulations accounting for protein interactions at atomic resolution are useful for characterizing the diffusion patterns in crowded environments. We present a comprehensive analysis of protein diffusion under different crowding conditions based on our recent docking-based approach simulating an intracellular crowded environment by sampling the intermolecular energy landscape using the Markov Chain Monte Carlo protocol. The procedure was extensively benchmarked, and the results are in very good agreement with the available experimental and theoretical data. The translational and rotational diffusion rates were determined for different types of proteins under crowding conditions in a broad range of concentrations. A protein system representing most abundant protein types in the E. coli cytoplasm was simulated, as well as large systems of other proteins of varying sizes in heterogeneous and self-crowding solutions. Dynamics of individual proteins was analyzed as a function of concentration and different diffusion rates in homogeneous and heterogeneous crowding. Smaller proteins diffused faster in heterogeneous crowding of larger molecules, compared to their diffusion in the self-crowded solution. Larger proteins displayed the opposite behavior, diffusing faster in the self-crowded solution. The results show the predictive power of our structure-based simulation approach for long timescales of cell-size systems at atomic resolution. 
    more » « less
  • Citation Formats
  • MLA
  • APA
  • Chicago
  • BibTeX
  • Save / Share this Record
  • Send to Email