Plastoglobule Lipid Droplet Isolation from Plant Leaf Tissue and Cyanobacteria

Shivaiah, Kiran‐Kumar; Susanto, Febri Adi; Devadasu, Elsinraju; Lundquist, Peter K.

doi:10.3791/64515

Cited by 3 publications

(2 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…From N. benthamiana plants expressing AtFBN1a, AtFBN5-A, and AtFBN7a variants, plastoglobules and thylakoid membranes were extracted according to Shivaiah et al, 2022 (90). Briefly, plastoglobules were separated using 0.2 M sucrose in Medium R solution (50 mM HEPES-KOH pH 8.0 and 5 mM MgCl2 with phosphatase and protease inhibitors) after sonicating thylakoid fractions.…”

Section: Methodsmentioning

confidence: 99%

An amphipathic helix drives interaction of Fibrillins with plastoglobule lipid droplets

Shivaiah,

Boren,

Tequia-Herrera

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Plastoglobule lipid droplets of chloroplasts serve complex roles affecting plant development, stress tolerance and photosynthesis. They harbor a set of approximately 42 proteins that collectively dictate plastoglobule functions. Due to the monolayer structure of plastoglobules which encompass a neutral lipid core, these proteins must associate monotopically on the plastoglobule surface. However, targeting determinants have not been identified for plastoglobule proteins, and the protein-membrane interaction mechanisms that establish the plastoglobule proteome remain unclear. Here, we demonstrate that plastoglobule-localized Fibrillins harbor an amphipathic helix at the lip of their β-barrel that is necessary for proper plastoglobule association. Molecular dynamics simulations support the specific interaction of the amphipathic helix of AtFBN1a with membranes rich in lipid packing defects which are expected to be especially prevalent on the tightly curved surface of plastoglobules. Introduction of one of the amphipathic helices into stromal-or thylakoid-localized FBNs was ineffective at redirecting the proteins to plastoglobules, likely due to endogenous protein-protein interactions that override the influence of the amphipathic helix. Proteomic analyses indicate AtFBN1a influences the plastoglobule proteome through outcompeting and recruiting specific proteins. We also demonstrate that the plastoglobule-localized FBNs, AtFBN1a and AtFBN7a, bind unsaturated fatty acids, particularly C18:1, and that elimination of the amphipathic helix suppresses fatty acid binding in AtFBN1a, but promotes fatty acid binding in AtFBN7a. Predicted amphipathic helices can be identified on two-thirds of plastoglobule proteins, indicating the use of amphipathic helices may be a general mechanism by which proteins selectively associate with plastoglobules.

show abstract

Section: Methodsmentioning

confidence: 99%

An amphipathic helix drives interaction of Fibrillins with plastoglobule lipid droplets

Shivaiah,

Boren,

Tequia-Herrera

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…They contain mainly prenylquinones and carotenoids and lower amounts of TAGs [128,129]. A protocol for their isolation has been proposed in 2022 that may lift a hurdle for their molecular characterisation [130].…”

Section: Other Chloroplast Microcompartmentsmentioning

confidence: 99%

Location of the photosynthetic carbon metabolism in microcompartments and separated phases in microalgal cells

Launay,

Avilan,

Gérard

et al. 2023

FEBS Letters

View full text Add to dashboard Cite

Carbon acquisition, assimilation and storage in eukaryotic microalgae and cyanobacteria occur in multiple compartments that have been characterised by the location of the enzymes involved in these functions. These compartments can be delimited by bilayer membranes, such as the chloroplast, the lumen, the peroxisome, the mitochondria or monolayer membranes, such as lipid droplets or plastoglobules. They can also originate from liquid–liquid phase separation such as the pyrenoid. Multiple exchanges exist between the intracellular microcompartments, and these are reviewed for the CO2 concentration mechanism, the Calvin–Benson–Bassham cycle, the lipid metabolism and the cellular energetic balance. Progress in microscopy and spectroscopic methods opens new perspectives to characterise the molecular consequences of the location of the proteins involved, including intrinsically disordered proteins.

show abstract

Role of isotropic lipid phase in the fusion of photosystem II membranes

Böde,

Javornik,

Dlouhý

et al. 2024

Photosynth Res

View full text Add to dashboard Cite

It has been thoroughly documented, by using 31P-NMR spectroscopy, that plant thylakoid membranes (TMs), in addition to the bilayer (or lamellar, L) phase, contain at least two isotropic (I) lipid phases and an inverted hexagonal (HII) phase. However, our knowledge concerning the structural and functional roles of the non-bilayer phases is still rudimentary. The objective of the present study is to elucidate the origin of I phases which have been hypothesized to arise, in part, from the fusion of TMs (Garab et al. 2022 Progr Lipid Res 101,163). We take advantage of the selectivity of wheat germ lipase (WGL) in eliminating the I phases of TMs (Dlouhý et al. 2022 Cells 11: 2681), and the tendency of the so-called BBY particles, stacked photosystem II (PSII) enriched membrane pairs of 300–500 nm in diameter, to form large laterally fused sheets (Dunahay et al. 1984 BBA 764: 179). Our 31P-NMR spectroscopy data show that BBY membranes contain L and I phases. Similar to TMs, WGL selectively eliminated the I phases, which at the same time exerted no effect on the molecular organization and functional activity of PSII membranes. As revealed by sucrose-density centrifugation, magnetic linear dichroism spectroscopy and scanning electron microscopy, WGL disassembled the large laterally fused sheets. These data provide direct experimental evidence on the involvement of I phase(s) in the fusion of stacked PSII membrane pairs, and strongly suggest the role of non-bilayer lipids in the self-assembly of the TM system.

show abstract

Plastoglobule Lipid Droplet Isolation from Plant Leaf Tissue and Cyanobacteria

Cited by 3 publications

References 0 publications

An amphipathic helix drives interaction of Fibrillins with plastoglobule lipid droplets

An amphipathic helix drives interaction of Fibrillins with plastoglobule lipid droplets

Location of the photosynthetic carbon metabolism in microcompartments and separated phases in microalgal cells

Role of isotropic lipid phase in the fusion of photosystem II membranes

Contact Info

Product

Resources

About