Translocation of cell-penetrating peptides involving calcium-dependent interactions between anionic glycosaminoglycans and phosphocholine bilayer DOI
Bingwei He, Sonia Khemaissa,

Sébastien Cardon

et al.

Published: Oct. 17, 2024

Cell-penetrating peptides can internalize ubiquitously in many, if not all, cell types. To explore the specific targeting issue of cell-penetrating (CPPs), we studied glycosaminoglycan (GAG)-binding previously identified Otx2 and En2 homeoproteins (HPs), alone or extended with penetratin-like third helix (H3) En2. HPs are indeed known to cells, thanks their GAG-targeting sequence (Joliot et al. 2022; Cardon 2023). We quantified capacity these enter into various lines express different levels types heparan sulfates (HS) chondroitin (CS) GAGs. also analyzed by calorimetry (DSC, ITC) fluorescence spectroscopy, binary ternary interactions between heparin (HI), (4S, 6S)CS (CS-E), zwitterionic phosphocholine (PC) model membranes those peptides. Altogether, our results demonstrate existence Ca2+-dependent CS-E HI PC lipid bilayers, major phospholipid found animal plasma membrane. Importantly, show that act as a bridge be exploited chimeric CS-E-recognition motif-H3 peptide bind cross membrane bilayer get access directly cytosol cells. this study brings further information uncovering molecular mechanism translocation process CPPs implies GAGs at cell-surface. It shed light on role paracrine activity specificity HPs.

Language: Английский

Binding affinities for 2D protein dimerization benefit from enthalpic stabilization DOI Creative Commons

Adip Jhaveri,

S Chhibber,

Naveen V. Kulkarni

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 21, 2025

Abstract Dimerization underpins all macromolecular assembly processes both on and off the membrane. While strength of dimerization, K D , is commonly quantified in solution (3D), many proteins like soluble BAR domain-containing also reversibly dimerize while bound to a membrane surface (2D). The ratio dissociation constants, defines lengthscale that essential for determining whether dimerization more favorable or surface, particularly these transition between 3D 2D. purely entropic rigid-body estimates h apply well transmembrane adhesion proteins, we show here using Molecular Dynamics simulations even moderate flexibility domains dramatically alters free energy landscape from 2D, driving enhanced selectivity stability native dimer By simulating homodimerization three distinct environments, 1) 2) lipid bilayer (2D), 3) fully solvated but restrained pseudo 2D environments induce backbone configurations better match crystal structure produce enthalpically states, violating drive ≪ RIGID . Remarkably, contact with an explicit not necessary changes, as induces this same result. We outcome depends protein interaction, parameterization produces exceptionally stable binding does systematic improvements With lengthscales calculated are below physiological volume-to-surface-area lengthscale, will be membrane, which aligns domain function remodelers. Our approach provides simple metrics move beyond affinities assess conformational selects membranes.

Language: Английский

Citations

0
Dynamic Plasma Membrane Topography Linked With Arp2/3 Actin Network Induction During Cell Shape Change DOI Creative Commons
Tony Harris

BioEssays, Journal Year: 2025, Volume and Issue: unknown

Published: March 31, 2025

ABSTRACT Recent studies show the importance of mesoscale changes to plasma membrane (PM) topography during cell shape change. Local folding and flattening surface is mechanosensitive, changing in response both microenvironment structural elements intracellular cytoskeletal activities. These elicit local mechanical signaling events that act conjunction with molecular signal transduction pathways remodel cortex. Experimental manipulations PM curvature its sufficiency for recruiting Arp2/3 actin network induction pathways. Additionally, diverse changes—ranging from neutrophil migration early Drosophila embryo cleavage neural stem asymmetric division—show generation linked induction, which then remodels dynamic control structure. examples are reviewed detail, together known potential causes changes, downstream effects, higher‐order feedback.

Language: Английский

Citations

0
Engineered Proteins and Chemical Tools to Probe the Cell Surface Proteome DOI Creative Commons
Kevin K. Leung, Kaitlin Schaefer, Zhi Lin

et al.

Chemical Reviews, Journal Year: 2025, Volume and Issue: unknown

Published: April 3, 2025

The cell surface proteome, or surfaceome, is the hub for cells to interact and communicate with outside world. Many disease-associated changes are hard-wired within yet approved drugs target less than 50 proteins. In past decade, proteomics community has made significant strides in developing new technologies tailored studying surfaceome all its complexity. this review, we first dive into unique characteristics functions of emphasizing necessity specialized labeling, enrichment, proteomic approaches. An overview surfaceomics methods provided, detailing techniques measure protein expression how leads novel discovery. Next, highlight advances proximity labeling (PLP), showcasing various enzymatic photoaffinity can map protein-protein interactions membrane complexes on surface. We then review role extracellular post-translational modifications, focusing glycosylation, proteolytic remodeling, secretome. Finally, discuss identifying tumor-specific peptide MHC they have shaped therapeutic development. This emerging field neo-protein epitopes constantly evolving, where targets identified at proteome level encompass defined PTMs, complexes, dysregulated cellular tissue locations. Given functional importance biology therapy, view as a critical piece quest neo-epitope

Language: Английский

Citations

0
A novel motif in calcimembrin/C16orf74 dictates multimeric dephosphorylation by calcineurin DOI Open Access

Devin A. Bradburn,

Joana Reis, Shariq Qayyum

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: May 13, 2024

Abstract Calcineurin, the Ca 2+ /calmodulin-activated protein phosphatase, recognizes substrates and regulators via short linear motifs, PxIxIT LxVP, which dock to distinct sites on calcineurin determine distribution catalysis, respectively. Calcimembrin/C16orf74 (CLMB), an intrinsically disordered microprotein whose expression correlates with poor cancer outcomes, targets membranes where it may promote oncogenesis by shaping signaling. We show that CLMB associates lipidation, i.e. N-myristoylation reversible S-acylation. Furthermore, contains unusual composite ‘LxVPxIxIT’ motif, binds PxIxIT-docking site extraordinarily high affinity when phosphorylated, 33 LxVPxIxITxx(p)T 44 . Calcineurin dephosphorylates decrease this affinity, but Thr44 is protected from dephosphorylation PxIxIT-bound. propose dephosphorylated in multimeric complexes, one PxIxIT-bound recruits membranes, allowing a second engage its LxVP motif be dephosphorylated. In vivo vitro data, including nuclear magnetic resonance (NMR) analyses of CLMB-calcineurin supports model. Thus, imposes unique properties signaling at sensitivity CLMB:calcineurin ratios, phosphorylation dynamic

Language: Английский

Citations

1
Searching Through Cellular Landscapes DOI
Elena F. Koslover

Published: Jan. 1, 2024

Language: Английский

Citations

0
Enzymatic reactions dictated by the 2D membrane environment DOI Open Access
Rui Bai, Chun‐Wei Lin

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Aug. 23, 2024

Abstract The cell membrane is a fundamental component of cellular architecture. Beyond serving as physical barrier that encloses the cytosol, it also provides crucial platform for numerous biochemical reactions. Due to unique two-dimensional and fluidic environment membrane, reactions occur on its surface are subject specific constraints. However, advantages disadvantages membrane-mediated have yet be thoroughly explored. In this study, we reconstitute classic proteolytic cleavage reaction at interface, designed real-time, single-molecule kinetic analysis. interactions between enzyme substrate near examined under different scenarios. Our findings reveal while significantly enhances enzymatic activity, imposes diffusion limitations reduce activity over time. By adjusting enzyme’s affinity an intermediate level, enable "hop" surface, overcoming these constraints sustaining high with faster kinetics. These results provide critical insights into role in regulating can broadly applied other membrane-associated interactions.

Language: Английский

Citations

0
Translocation of cell-penetrating peptides involving calcium-dependent interactions between anionic glycosaminoglycans and phosphocholine bilayer DOI
Bingwei He, Sonia Khemaissa,

Sébastien Cardon

et al.

Published: Oct. 17, 2024

Cell-penetrating peptides can internalize ubiquitously in many, if not all, cell types. To explore the specific targeting issue of cell-penetrating (CPPs), we studied glycosaminoglycan (GAG)-binding previously identified Otx2 and En2 homeoproteins (HPs), alone or extended with penetratin-like third helix (H3) En2. HPs are indeed known to cells, thanks their GAG-targeting sequence (Joliot et al. 2022; Cardon 2023). We quantified capacity these enter into various lines express different levels types heparan sulfates (HS) chondroitin (CS) GAGs. also analyzed by calorimetry (DSC, ITC) fluorescence spectroscopy, binary ternary interactions between heparin (HI), (4S, 6S)CS (CS-E), zwitterionic phosphocholine (PC) model membranes those peptides. Altogether, our results demonstrate existence Ca2+-dependent CS-E HI PC lipid bilayers, major phospholipid found animal plasma membrane. Importantly, show that act as a bridge be exploited chimeric CS-E-recognition motif-H3 peptide bind cross membrane bilayer get access directly cytosol cells. this study brings further information uncovering molecular mechanism translocation process CPPs implies GAGs at cell-surface. It shed light on role paracrine activity specificity HPs.

Language: Английский

Citations

0