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Protocol for Isolation of Cardiomyocyte from Adult Mouse and Rat

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Femtoliter Injection of ESCRT-III Proteins into Adhered Giant Unilamellar Vesicles

Authors: Vasil N. Georgiev, Yunuen Avalos-Padilla, Xavier Fernàndez-Busquets and Rumiana Dimova, date: 02/20/2022, view: 1402, Q&A: 0

[Abstract]

The endosomal sorting complex required for transport (ESCRT) machinery mediates membrane fission reactions that exhibit a different topology from that observed in clathrin-coated vesicles. In all of the ESCRT-mediated events, the nascent vesicle buds away from the cytosol. However, ESCRT proteins are able to act upon membranes with different

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Activity-based Crosslinking to Identify Substrates of Thioredoxin-domain Proteins in Malaria Parasites

Authors: David W. Cobb, Grace S. Woods and Vasant Muralidharan, date: 02/20/2022, view: 1058, Q&A: 0

[Abstract] Malaria remains a major public health issue, infecting nearly 220 million people every year. The spread of drug-resistant strains of Plasmodium falciparum around the world threatens the progress made against this disease. Therefore, identifying druggable and essential pathways in P. falciparum parasites remains a major area of ...

Sex-specific Separation of Plasmodium falciparum Gametocyte Populations

Authors: Melanie C. Ridgway, Daniela Cihalova and Alexander G. Maier, date: 06/05/2021, view: 2016, Q&A: 0

[Abstract]

Plasmodium falciparum is a unicellular eukaryotic parasite that causes malaria in humans. The parasite is spread by Anopheles mosquitoes after ingestion of sexual stage parasites known as gametocytes. Malaria transmission depends on parasites switching from the disease-causing asexual blood forms to male and female gametocytes. The current

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MicroScale Thermophoresis as a Tool to Study Protein-peptide Interactions in the Context of Large Eukaryotic Protein Complexes

Authors: Maximilian G. Plach, Klaus Grasser and Thomas Schubert, date: 12/05/2017, view: 20317, Q&A: 0

[Abstract] Protein-peptide interactions are part of many physiological processes, for example, epigenetics where peptide regions of histone complexes are crucial for regulation of chromatin structure. Short peptides are often also used as alternatives to small molecule drugs to target protein complexes. Studying the interactions between proteins and peptides ...

Liposome Disruption Assay to Examine Lytic Properties of Biomolecules

Authors: John R. Jimah, Paul H. Schlesinger and Niraj H. Tolia, date: 08/05/2017, view: 11435, Q&A: 0

[Abstract] Proteins may have three dimensional structural or amino acid features that suggest a role in targeting and disrupting lipids within cell membranes. It is often necessary to experimentally investigate if these proteins and biomolecules are able to disrupt membranes in order to conclusively characterize the function of these biomolecules. Here, we ...

A Gas Chromatography-Mass Spectrometry-Based Two Stage Assay for Measurement of in vitro myo-Inositol 3-phosphate Synthase (INO1) Activity

Authors: James I. MacRae and Malcolm J. McConville, date: 03/05/2015, view: 8888, Q&A: 0

[Abstract] This method describes an in vitro assay for measuring INO1 enzyme activity (the conversion of glucose 6-phosphate to myo-inositol 3-phosphate) in cell-free extracts. The method was first described for Plasmodium falciparum cells in MacRae et al. (2014) and consists of two parts: Part 1 describes the assay itself ...

Preparation of Parasite Protein Extracts and Western Blot Analysis

Authors: Arlett Heiber and Tobias Spielmann, date: 06/05/2014, view: 22518, Q&A: 0

[Abstract] In order to prepare protein extracts of Plasmodium falciparum blood stages for western blot analysis, infected red blood cells (iRBC) need to be separated from uninfected red blood cells (uRBC) which make up the bulk of the parasite culture. Depending on the localisation of the parasite protein of interest, different methods are available ...

Cyclic Nucleotide (cAMP and cGMP) Assays and Capture ELISA for Quantitative Analysis of Plasmodium falciparum Blood-stage Egress

Authors: Fiona Hackett, Christine R Collins, Malcolm Strath and Michael J Blackman, date: 03/05/2014, view: 9615, Q&A: 0

[Abstract] Upon rupture of Plasmodium falciparum (P. falciparum) schizonts in vitro (an event known as egress), merozoites are released into the culture medium. The merozoites invade fresh red blood cells, a process that involves shedding of a microneme protein called apical membrane antigen-1 (AMA1) from the merozoite surface. ...

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