Light-Inducible Generation of Membrane Curvature in Live Cells with Engineered Bar Domain Proteins

Abstract

to mimic the presence of a negatively charged phosphoryl group. This is useful since intact phosphorylated Cc is difficult to purify by traditional methods. The binding affinity that the mutant Cc has for wild-type CcO and the electron transfer rates between the mutant Cc and wild-type CcO will be determined. Ultimately, this project will provide insight that is needed to settle the current debate over the binding model for Cc and CcO. There are currently two main models proposed; however, they each vary in the role of Cc's S47 residue during Cc and CcO binding. Supported by NIH grants GM20488 and 8P30GM103450.

Chiral growth of adherent filopodia

“…As per the second question, namely how can such shift trigger a leftward turning of the filopodia, several general mechanisms might be considered. Recent published data suggests that filopodia growth is initiated by membrane remodeling with inverted BAR-domain proteins (Jones et al, 2020), while consequent polymerization of actin filaments stabilizes the protrusions and promotes their elongation.…”

“…As per the second question, namely how can such shift trigger a leftward turning of the filopodia, several general mechanisms might be considered. Recent published data suggests that filopodia growth is initiated by membrane remodeling with inverted BAR-domain proteins (Jones et al, 2020), while consequent polymerization of actin filaments stabilizes the protrusions and promotes their elongation. Assuming the concentration of the I-BAR domain proteins to be homogeneous all over the filopodial membrane, the mean membrane curvature (½ (c 1 +c 2 ) = ½ (1/r 1 +1/r 2 ), where c 1 and c 2 are the principal curvatures, and r 1 and r 2 the principal radii of curvature, respectively) should be constant over the entire filopodia surface.…”

CRY-BARs: Versatile light-gated molecular tools for the remodeling of membrane architectures

“…In recent years, the critical role of proteins involved in membrane curvature inducing and sensing has come to light (Antonny, 2011). In particular, BAR domain containing proteins possess diverse activities at the plasma membrane (Linkner et al, 2014; Prévost et al, 2015; Pykäläinen et al, 2011; Saarikangas et al, 2015, 2011; Yu et al, 2011; Zhao, Pykäläinen, & Lappalainen, 2011); as such, a heightened understanding of these roles has invited recent investigations of their suitability for control with optogenetic tools (Jones, Liu, & Cui, 2020). In this work, we investigate the potential of an inverse BAR (I-BAR) domain from the MTSS1 protein (Missing in Metastasis 1) to serve as an actuator of membrane architecture and plasma membrane dynamics.…”

A study of lipid-mediated clustering of oncogenic Ras proteins on cell membrane using nanotopography engineering

“…138 By incorporating optogenetic control into the curvature-generating proteins, this method is more accurate, reversible, and controllable in location and the time point to switch on and off the event. 139 Hypotonic treatment acts as a way to induce flatter membranes for comparison. 138 Besides the artificially produced curved membrane structures, the protein enrichment in filopodia, if related, can also be measured based on the fluorescence-labelled PM signal to compute the curvature-dependent protein gathering.…”