Screening for and characterization of phospholipase A 1 hypersecretory mutants of Tetrahymena thermophila

Hartmann, Marcus W W; Guberman, A.; Florín-Christensen, Mónica; Tiedtke, Arno

doi:10.1007/s002530000405

Cited by 24 publications

(12 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Several of the identified Tetrahymena proteins fall in this category, including proteins with strong matches to cathepsins L and B, acid alpha-glucosidase, and acid phosphatases. Also included in this category is lysosomal phospholipase A 1 , which has previously been shown to be secreted by Tetrahymena (38). Since it is unlikely that the phagosome preparation procedures we employed would have resulted in the recovery of proteins secreted into the culture medium, the detection of this hydrolytic enzyme in our analysis strongly suggests that it is also localized to phagosomes.…”

Section: Vol 5 2006mentioning

confidence: 89%

The Tetrahymena thermophila Phagosome Proteome

et al. 2006

View full text Add to dashboard Cite

In vertebrates, phagocytosis occurs mainly in specialized cells of the immune system and serves as a primary defense against invading pathogens, but it also plays a role in clearing apoptotic cells and in tissue remodeling during development. In contrast, unicellular eukaryotes, such as the ciliate Tetrahymena thermophila, employ phagocytosis to ingest and degrade other microorganisms to meet their nutritional needs. To learn more about the protein components of the multistep process of phagocytosis, we carried out an analysis of the Tetrahymena phagosome proteome. Tetrahymena cells were fed polystyrene beads, which allowed for the efficient purification of phagosomes. The protein composition of purified phagosomes was then analyzed by multidimensional separation coupled with tandem mass spectrometry. A total of 453 peptides were identified that resulted in the identification of 73 putative phagosome proteins. Twenty-eight of the proteins have been implicated in phagocytosis in other organisms, indicating that key aspects of phagocytosis were conserved during evolution. Other identified proteins have not previously been associated with phagocytosis, including some of unknown function. Live-cell confocal fluorescence imaging of Tetrahymena strains expressing green fluorescent protein-tagged versions of four of the identified phagosome proteins provided evidence that at least three of the proteins (including two with unknown functions) are associated with phagosomes, indicating that the bulk of the proteins identified in the analyses are indeed phagosome associated.

show abstract

Section: Vol 5 2006mentioning

confidence: 89%

The Tetrahymena thermophila Phagosome Proteome

et al. 2006

View full text Add to dashboard Cite

show abstract

“…Part of the interest in characterizing secretion comes from the question of whether Tetrahymena can be usefully engineered to produce and secrete heterologous proteins (Aldag et al, 2011; Hartmann et al, 2000). …”

Section: Recent Work On Membrane Traffic In Tetrahymenamentioning

confidence: 99%

Conservation and Innovation in Tetrahymena Membrane Traffic: Proteins, Lipids, and Compartments

Nusblat

Bright

Turkewitz

2012

Methods in Cell Biology

View full text Add to dashboard Cite

“…generally secreted a number of lysosomal enzymes, including PLA 2 , PLC, PLD, and lipase. PLA 1 -hypersecretory mutants of T. thermophila were isolated using random chemical mutagenesis (34), and a practical purification method was designed for large-scale production of the PLA 1 enzyme (35). The purification process, based on the selective interaction of T. thermophila PLA 1 with its liposomal substrate, was similar to affinity chromatography but less expensive, as no substrate immobilization was required.…”

Section: Plamentioning

confidence: 99%

Phospholipases: Occurrence and production in microorganisms, assay for high‐throughput screening, and gene discovery from natural and man‐made diversity

Song

Han

Rhee

2005

J Americ Oil Chem Soc

View full text Add to dashboard Cite

Various kinds of phospholipids have wide industrial applications such as in food and nutraceuticals, cosmetics, agricultural products, and pharmaceuticals. The demand for reliable biocatalysts for the production of phospholipid products, such as phospholipases A 1 , A 2 , C, and D, has steadily increased over the past several decades. A large number of microbial phospholipases have been isolated and characterized, and the increasing availability of these enzymes could eventually lead to the sustained development of phospholipid-related biotechnology. Although a number of reactions have been performed using phospholipases, a reliable and efficient supply of superior phospholipases in quantity is still a challenge for their practical application. High-throughput functional assay methods for phospholipases should be devised to develop superior new species from the huge diversity of phospholipases. Recent biotechnological advances in the discovery of new phospholipase genes from natural sources, such as extremophiles and metagenome libraries, and in the functional enhancement of phospholipases by protein engineering, such as directed evolution, can provide valuable means of rapidly developing practical uses of phospholipases for various applications.

show abstract

Screening for and characterization of phospholipase A 1 hypersecretory mutants of Tetrahymena thermophila

Cited by 24 publications

References 5 publications

The Tetrahymena thermophila Phagosome Proteome

The Tetrahymena thermophila Phagosome Proteome

Conservation and Innovation in Tetrahymena Membrane Traffic: Proteins, Lipids, and Compartments

Phospholipases: Occurrence and production in microorganisms, assay for high‐throughput screening, and gene discovery from natural and man‐made diversity

Contact Info

Product

Resources

About