Isolation of Lipid Droplets for Protein and Lipid Analysis

Horn, Patrick J.; Chapman, Kent D.; Ischebeck, Till

doi:10.1007/978-1-0716-1362-7_16

Cited by 6 publications

(2 citation statements)

References 60 publications

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“…Oleosin is associated with small oil bodies, whereas very large oil bodies lack oleosins and are stabilized by the lipid-associated proteins LDAP1 and LDAP2 ( Horn et al 2021 ). Interestingly, depending on the presence of oleosin, the oil bodies are variable in size.…”

Section: Discussionmentioning

confidence: 99%

Degeneration of oil bodies by rough endoplasmic reticulum (rER)-associated protein during seed germination in Cannabis sativa L.

Kim,

Han,

Olejar

et al. 2023

AoB PLANTS

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Abstract. Oil bodies serve as a vital energy source of embryos during germination and contribute to sustaining the initial growth of seedlings until photosynthesis initiation. Despite high stability in chemical properties, how oil bodies break down and go into the degradation process during germination is still unknown. This study provides a morphological understanding of the mobilization of stored compounds in the seed germination of Cannabis. The achenes of fibrous hemp cultivar (Cannabis sativa cv. ‘Chungsam’) were examined in this study using light microscopy, scanning electron microscopy and transmission electron microscopy. Oil bodies in Cannabis seeds appeared spherical and sporadically distributed in the cotyledonary cells. Protein bodies contained electron-dense globoid and heterogeneous protein matrices. During seed germination, rough endoplasmic reticulum (rER) and high electron-dense substances were present adjacent to the oil bodies. The border of the oil bodies became a dense cluster region and appeared as a sinuous outline. Later, irregular hyaline areas were distributed throughout oil bodies, showing the destabilized emulsification of oil bodies. Finally, the oil bodies lost their morphology and fused with each other. The storage proteins were concentrated in the centre of the protein body as a dense homogenous circular mass surrounded by a light heterogeneous area. Some storage proteins are considered emulsifying agents on the surface region of oil bodies, enabling them to remain stable and distinct within and outside cotyledon cells. At the early germination stage, rER appeared and dense substances aggregated adjacent to the oil bodies. Certain proteins were synthesized within the rER and then translocated into the oil bodies by crossing the half membrane of oil bodies. Our data suggest that rER-associated proteins function as enzymes to lyse the emulsifying proteins, thereby weakening the emulsifying agent on the surface of the oil bodies. This process plays a key role in the degeneration of oil bodies and induces coalescence during seed germination.

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Section: Discussionmentioning

confidence: 99%

Degeneration of oil bodies by rough endoplasmic reticulum (rER)-associated protein during seed germination in Cannabis sativa L.

Kim,

Han,

Olejar

et al. 2023

AoB PLANTS

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“…Proteomic RAW data were processed in MaxQuant 2.0.3.1 software (Cox & Mann, 2008; Tyanova, Temu, & Cox, 2016) on default settings except enabling “Label‐free quantification ‐ LFQ,” “iBAQ,” “FTMS recalibration,” “Match between runs,” and set “Intensity determination” to “Total sum.” The P. patens primary transcript protein file (v3.3, DOE‐JGI; https://phytozome-next.jgi.doe.gov/; Goodstein et al., 2012; Lang et al., 2018) was used as reference library. Filtering and relative iBAQ (riBAQ) normalization (‰ of total protein abundance per sample) from the resulting MaxQuant proteinGroups file was performed in Perseus 1.6.2.2 (Tyanova, Temu, Sinitcyn, et al., 2016) as previously described (Horn et al., 2021). The raw data on the N. tabacum pollen proteome (Ischebeck et al., 2014) was re‐processed with MaxQuant using the same settings and the database based on the N. tabacum genome (Edwards et al., 2017).…”

Section: Methodsmentioning

confidence: 99%

Proteome plasticity during Physcomitrium patens spore germination – from the desiccated phase to heterotrophic growth and reconstitution of photoautotrophy

Hembach,

Niemeyer,

Schmitt

et al. 2023

The Plant Journal

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SUMMARYThe establishment of moss spores is considered a milestone in plant evolution. They harbor protein networks underpinning desiccation tolerance and accumulation of storage compounds that can be found already in algae and that are also utilized in seeds and pollen. Furthermore, germinating spores must produce proteins that drive the transition through heterotrophic growth to the autotrophic plant. To get insight into the plasticity of this proteome, we investigated it at five timepoints of moss (Physcomitrium patens) spore germination and in protonemata and gametophores. The comparison to previously published Arabidopsis proteome data of seedling establishment showed that not only the proteomes of spores and seeds are functionally related, but also the proteomes of germinating spores and young seedlings. We observed similarities with regard to desiccation tolerance, lipid droplet proteome composition, control of dormancy, and β‐oxidation and the glyoxylate cycle. However, there were also striking differences. For example, spores lacked any obvious storage proteins. Furthermore, we did not detect homologs to the main triacylglycerol lipase in Arabidopsis seeds, SUGAR DEPENDENT1. Instead, we discovered a triacylglycerol lipase of the oil body lipase family and a lipoxygenase as being the overall most abundant proteins in spores. This finding indicates an alternative pathway for triacylglycerol degradation via oxylipin intermediates in the moss. The comparison of spores to Nicotiana tabacum pollen indicated similarities for example in regards to resistance to desiccation and hypoxia, but the overall developmental pattern did not align as in the case of seedling establishment and spore germination.

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Cellular organelles as drug carriers for disease treatment

Hu,

Liu,

et al. 2023

Journal of Controlled Release

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Isolation of Lipid Droplets for Protein and Lipid Analysis

Cited by 6 publications

References 60 publications

Degeneration of oil bodies by rough endoplasmic reticulum (rER)-associated protein during seed germination in Cannabis sativa L.

Degeneration of oil bodies by rough endoplasmic reticulum (rER)-associated protein during seed germination in Cannabis sativa L.

Proteome plasticity during Physcomitrium patens spore germination – from the desiccated phase to heterotrophic growth and reconstitution of photoautotrophy

Cellular organelles as drug carriers for disease treatment

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