ArticleAnalytical biochemistry2008
Real-time assay for monitoring membrane association of lipid-binding domains.
Article in Analytical biochemistry, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
11 citing papers in PubMed, 20 citations in OpenAlex.
- Membrane fission during bacterial spore development requires cellular inflation driven by DNA translocation.Current biology : CB · 2022Article
- FisB relies on homo-oligomerization and lipid binding to catalyze membrane fission in bacteria.PLoS biology · 2021Article
- Doc2b CaScientific reports · 2019Article
- Sphingomimetic multiple sclerosis drug FTY720 activates vesicular synaptobrevin and augments neuroendocrine secretion.Scientific reports · 2017Article
- Cellular and molecular interactions of phosphoinositides and peripheral proteins.Chemistry and physics of lipids · 2014Review
- Membrane-proximal tryptophans of synaptobrevin II stabilize priming of secretory vesicles.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2012Article
- Phospholipid association is essential for dynamin-related protein Mgm1 to function in mitochondrial membrane fusion.The Journal of biological chemistry · 2009Article
- Solution and membrane-bound conformations of the tandem C2A and C2B domains of synaptotagmin 1: Evidence for bilayer bridging.Journal of molecular biology · 2009Article
- Sphingosine facilitates SNARE complex assembly and activates synaptic vesicle exocytosis.Neuron · 2009Article
- Multiple RIBEYE-RIBEYE interactions create a dynamic scaffold for the formation of synaptic ribbons.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2008Article
- DOC2B acts as a calcium switch and enhances vesicle fusion.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2008Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
The C2 domain is a common protein module which mediates calcium-dependent phospholipid binding. Several assays have previously been developed to measure membrane association. However, these assays either have technical drawbacks or are laborious to carry out. We now present a simple solution-based turbidity method for rapidly assaying membrane association of single lipid-binding domains in real time. We used the first C2 domain of synaptotagmin 1 (C2A) as a model lipid-binding moiety. Our use of the common dimeric glutathione-S-transferase (GST) fusion tag allowed two C2A domains to be brought into close proximity. Consequently, calcium-triggered phospholipid binding by this artificially dimerized C2A resulted in liposomal aggregation, easily assayed by following absorbance of the solution at 350 nm. The assay is simple and sensitive and can be scaled up conveniently for use in a multiwell plate format, allowing high-throughput screening. In our screens, we identified nickel as a novel activator of synaptotagmin 1 C2A domain membrane association. Finally, we show that the turbidity method can be applied to the study of other GST-tagged lipid-binding proteins such as epsin, protein kinase C-beta, and synaptobrevin.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.