Experimentally determined leaflet-leaflet phase diagram of an asymmetric lipid bilayer

Autor(es)

Enoki, Thais A. ; Heberle, Frederick A.

• Tipo de documento: Artigo Científico
• Fonte: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA; v. 120, n. 46, p. 11-pg., 2023-11-14.

Resumo

We have determined the partial leaflet-leaflet phase diagram of an asymmetric lipid bilayer at ambient temperature using asymmetric giant unilamellar vesicles (aGUVs). Symmetric GUVs with varying amounts of 1,2- dipalmitoyl- sn- glycero-3- phosphocholine and DOPC (1,2- dioleoyl- sn- glycero-3- phosphocholine) were hemifused to a supported lipid bilayer (SLB) composed of DOPC, resulting in lipid exchange between their outer leaflets. The GUVs and SLB contained a red and green lipid fluorophore, respectively, thus enabling the use of confocal fluorescence imaging to determine both the extent of lipid exchange (quantified for individual vesicles by the loss of red intensity and gain of green intensity) and the presence or absence of phase separation in aGUVs. Consistent with previous reports, we found that hemifusion results in large variation in outer leaflet exchange for individual GUVs, which allowed us to interrogate the phase behavior at multiple points within the asymmetric composition space of the binary mixture. When initially symmetric GUVs showed coexisting gel and fluid domains, aGUVs with less than similar to 50% outer leaflet exchange were also phase- separated. In contrast, aGUVs with greater than 50% outer leaflet exchange were uniform and fluid. In some cases, we also observed three coexisting bilayer- spanning phases: two registered phases and an anti- registered phase. These results suggest that a relatively large unfavorable midplane interaction between ordered and disordered phases in opposing leaflets (i.e., a midplane surface tension) can overwhelm the driving force for lateral phase separation within one of the leaflets, resulting in an asymmetric bilayer with two uniformly mixed leaflets that is poised to phase- separate upon leaflet scrambling. (AU)

• Processo FAPESP: 23/05540-5 - Estudos da assimetria da membrana plasmática: membranas modelo mimetizando células saudáveis e cancerígenas, e estratégias terapêuticas para tratamento de câncer
• Beneficiário: Thais Azevedo Enoki Liarte
• Modalidade de apoio: Bolsas no Brasil - Jovens Pesquisadores
• Processo FAPESP: 22/04046-4 - Estudos da assimetria da membrana plasmática: membranas modelo mimetizando células saudáveis e cancerígenas, e estratégias terapêuticas para tratamento de câncer
• Beneficiário: Thais Azevedo Enoki Liarte
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores