Alkyl diazirines are photo-reactive functional groups frequently used in photo-affinity labeling (PAL). They are compact in size, being nearly isosteric to a methyl group, and they are activated at a wavelength of light (~355 nm) that is not damaging to protein(s). Photo-crosslinked protein targets are then visualized by the reporter group (e.g. fluorophore, biotin, or radioactive label). Covalent bond formation between the probe and targets enable the subsequent purification and identification of the targets using techniques such as SDS-PAGE, immunoprecipitation, biotin-streptavidin affinity purification and mass spectrometry. Alkyl diazirines are photo-stable under typical laboratory lighting conditions so there is no need to perform experiments in the dark.
New photocrosslinking compounds you will find here
The first rapidly reversible chemical dimerization system – rCD1 – , which permits to determine kinetics of lipid metabolizing enzymes in living cells, is now available at SiChem . This new system was applied to induce and stop the activity of phosphatidyl 3-kinase (PI3K) (SC-7000_Chemical_Dimerizer_info). rCD1 is available in 2 sizes: 100µg (for app. 20 experiments) and 500µg (SC-7000)
A representative time-lapse fluorescence movie of HeLa cells co-transfected with NLS2-SNAPf-ECFP and mRFP-FKBP constructs. rCD1 (1 μM) induces translocation of mRFP-FKBP into the nucleus; addition of FK506 (1 μM) leads to release of mRFP-FKBP from the nucleus !
INO-4995 (1-O-octyl-2-O-butyryl-D-myo-inositol 3,4,5,6-tetrakisphosphate octakis(propionoxymethyl)ester) is an ion channel modulator that is slowly metabolized in cells. Studies have shown that it inhibits ENaC activity while promoting expression of the calcium-activated chloride channel, TMEM16a, in membranes. Thus it has potential therapeutic activity in the treatment of cystic fibrosis where it normalized nasal potential difference (npd) in a murine CF knock-out model and promoted fluid secretion in human CF airway epithelia.
Strong preclinical efficacy of INO-4995: a membrane-permeant Inositol-phosphate derivative has potential in treatment of Cystic Fibrosis (CF)
This graph is showing the difference in npd between mice before and after 4-day treatment with INO-4995 (0.0008mg/kg):
Am J Respir Cell Mol Biol 2010, 42(1), 105
The competition ” 100 Landmarks in the Land of Ideas” promotes ideas for urgent issues of tomorrow. The i3 – Life Sciences Cluster tries to find answers to the increasing life expectancy and the associated diseases. Innovative diagnostic companies in the Bremen / Bremerhaven region have joined forces to investigate biomarkers that may indicate certain diseases more accurately.
Fotograf. G. Ketschau (BIS, Bremerhaven)
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