Identification of the Polyhydroxybutyrate Granules in Mammalian Cultured Cells

Elustondo, Pia A.; Zakharian, Eleonora; Pavlov, Evgeny

doi:10.1002/cbdv.201200294

Cited by 15 publications

(12 citation statements)

References 21 publications

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“…However, structures resembling PHB granules were recently detected in human cells (Elustondo et al, 2012). These structures could be stained with lipophilic BODIPY R dye (a fluorescent boron dipyrromethene derivative) and were sensitive to treatment with PHB depolymerase PhaZ7 of Paucimonas lemoignei but insensitive to a catalytically inactive PhaZ7 variant.…”

Section: Storage Phbmentioning

confidence: 99%

New insights in the formation of polyhydroxyalkanoate granules (carbonosomes) and novel functions of poly(3‐hydroxybutyrate)

Jendrossek

Pfeiffer

2014

Environmental Microbiology

212

179

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SummaryThe metabolism of polyhydroxybutyrate (PHB) and related polyhydroxyalkanoates (PHAs) has been investigated by many groups for about three decades, and good progress was obtained in understanding the mechanisms of biosynthesis and biodegradation of this class of storage molecules. However, the molecular events that happen at the onset of PHB synthesis and the details of the initiation of PHB/PHA granule formation, as well as the complex composition of the proteinaceous surface layer of PHB/PHA granules, have only recently come into the focus of research and were not reviewed yet. In this contribution, we summarize the progress in understanding the initiation and formation of the PHA granule complex at the example of Ralstonia eutropha H16 (model organism of PHB-accumulating bacteria). Where appropriate, we include information on PHA granules of Pseudomonas putida as a representative species for medium-chain-length PHA-accumulating bacteria. We suggest to replace the previous micelle mode of PHB granule formation by the Scaffold Model in which the PHB synthase initiation complex is bound to the bacterial nucleoid. In the second part, we highlight data on other forms of PHB: oligo-PHB with ≈100 to 200 3-hydroxybutyrate (3HB) units and covalently bound PHB (cPHB) are unrelated in function to storage PHB but are presumably present in all living organisms, and therefore must be of fundamental importance.

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Section: Storage Phbmentioning

confidence: 99%

New insights in the formation of polyhydroxyalkanoate granules (carbonosomes) and novel functions of poly(3‐hydroxybutyrate)

Jendrossek

Pfeiffer

2014

Environmental Microbiology

212

179

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“…It is increasingly apparent that cPHB-proteins are ubiquitous in biological cells, both prokaryotic and eukaryotic [8,11–14,28,42–44]. However, cPHB has remained largely unnoticed because, unlike the highly-visible phase-bright PHB granules, cPHB is sparse and obscure.…”

Section: Cphb-proteinsmentioning

confidence: 99%

“…The granules have attained considerable commercial importance as ingredients of biodegradable plastics. Recently, Elustondo et al [8], using fluorescence spectroscopy, identified PHB-rich granules, which resembled bacterial carbanosomes and did not co-localize with mitochondria, lysosomes or endoplasmic reticulum, in the cytoplasm of cultured mammalian cells. It was suggested that the PHB may serve as a carbon and energy store for these cells.…”

Section: Introductionmentioning

confidence: 99%

The Role of Short-Chain Conjugated Poly-(R)-3-Hydroxybutyrate (cPHB) in Protein Folding

Reusch

2013

IJMS

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Poly-(R)-3-hydroxybutyrate (PHB), a linear polymer of R-3-hydroxybutyrate (R-3HB), is a fundamental constituent of biological cells. Certain prokaryotes accumulate PHB of very high molecular weight (10,000 to >1,000,000 residues), which is segregated within granular deposits in the cytoplasm; however, all prokaryotes and all eukaryotes synthesize PHB of medium-chain length (~100–200 residues) which resides within lipid bilayers or lipid vesicles, and PHB of short-chain length (<12 residues) which is conjugated to proteins (cPHB), primarily proteins in membranes and organelles. The physical properties of cPHB indicate it plays important roles in the targeting and folding of cPHB-proteins. Here we review the occurrence, physical properties and molecular characteristics of cPHB, and discuss its influence on the folding and structure of outer membrane protein A (OmpA) of Escherichia coli.

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“…A previous study has suggested that PHB is associated in granules that accumulate in the cytoplasm, similar to the behavior in bacteria, which might constitute an ‘energy reservoir’. In the same study, the colocalization of PHV granules with organelles was inspected in U87 cells, showing no remarkable colocalization with lysosomes, mitochondria, or endoplasmic reticulum [17]. …”

Section: Introductionmentioning

confidence: 99%

Intracellular trafficking and cellular uptake mechanism of PHBV nanoparticles for targeted delivery in epithelial cell lines

et al. 2017

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BackgroundNanotechnology is a science that involves imaging, measurement, modeling and a manipulation of matter at the nanometric scale. One application of this technology is drug delivery systems based on nanoparticles obtained from natural or synthetic sources. An example of these systems is synthetized from poly(3-hydroxybutyrate-co-3-hydroxyvalerate), which is a biodegradable, biocompatible and a low production cost polymer. The aim of this work was to investigate the uptake mechanism of PHBV nanoparticles in two different epithelial cell lines (HeLa and SKOV-3).ResultsAs a first step, we characterized size, shape and surface charge of nanoparticles using dynamic light scattering and transmission electron microscopy. Intracellular incorporation was evaluated through flow cytometry and fluorescence microscopy using intracellular markers. We concluded that cellular uptake mechanism is carried out in a time, concentration and energy dependent way. Our results showed that nanoparticle uptake displays a cell-specific pattern, since we have observed different colocalization in two different cell lines. In HeLa (Cervical cancer cells) this process may occur via classical endocytosis pathway and some internalization via caveolin-dependent was also observed, whereas in SKOV-3 (Ovarian cancer cells) these patterns were not observed. Rearrangement of actin filaments showed differential nanoparticle internalization patterns for HeLa and SKOV-3. Additionally, final fate of nanoparticles was also determined, showing that in both cell lines, nanoparticles ended up in lysosomes but at different times, where they are finally degraded, thereby releasing their contents.ConclusionsOur results, provide novel insight about PHBV nanoparticles internalization suggesting that for develop a proper drug delivery system is critical understand the uptake mechanism.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-016-0241-6) contains supplementary material, which is available to authorized users.

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Identification of the Polyhydroxybutyrate Granules in Mammalian Cultured Cells

Cited by 15 publications

References 21 publications

New insights in the formation of polyhydroxyalkanoate granules (carbonosomes) and novel functions of poly(3‐hydroxybutyrate)

New insights in the formation of polyhydroxyalkanoate granules (carbonosomes) and novel functions of poly(3‐hydroxybutyrate)

The Role of Short-Chain Conjugated Poly-(R)-3-Hydroxybutyrate (cPHB) in Protein Folding

Intracellular trafficking and cellular uptake mechanism of PHBV nanoparticles for targeted delivery in epithelial cell lines

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