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Description
FtsZ is a protein involved in bacterial cell division. This study examines the behavior of FtsZ under two-dimensional confinement and lateral spatial restrictions as an alternative to its three-dimensional confinement within a bacterium. Two protocols were developed to restrict the lateral movement of the protein in bilayers containing a lipid mixture of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) with 5% Nitrilotriacetic acid (NTA).
The first protocol involves creating micrometer-sized corrals using bovine serum albumin (BSA) / Polyethylene Glycol (PEG)-impregnated seals on pre-cleaned glass, then introducing the bilayer into these corrals and adding the FtsZ protein. The second protocol uses bilayer islands generated from a solution of Giant Unilamellar Vesicles (GUVs) at very low concentration on an atomically flat mica surface.
Once FtsZ is confined in this two-dimensional setup, its behavior was studied using Atomic Force Microscopy (AFM). The results represent an advance in FtsZ confinement techniques, although significant challenges were encountered, particularly with the first method. The confinement allowed for observations of distinct behaviors in the folding and distribution of FtsZ, suggesting its ability to modify and generate NTA patches in the fluid bilayer. These findings open new possibilities for FtsZ confinement methods and the development of protocols to enhance understanding of its behavior in the bacterial membrane, contributing to insights into bacterial division mechanisms and potential antibiotic design.
