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Single-Protein Nanopore Method Detects Post-Translational Modifications

At the University of Oxford, scientists have developed a nanopore technology that can identify three different post-translational modifications (PTMs) in individual proteins, even deep within long protein chains. The scientists asserted that their technology “[lays] the groundwork for compiling inventories of the proteoforms in cells and tissues.”

The technology was introduced in Nature Nanotechnology, in a paper titled, “Enzyme-less nanopore detection of post-translational modifications within long polypeptides.” The paper noted that single-molecule proteoform identification requires knowledge of the architecture of long polypeptide chains, knowledge that has proven elusive. Although there are methods for translocating folded proteins through solid-state nanopores or protein nanopores of large sizes, these methods have yet to locate PTMs within a polypeptide sequence. The methods that have detected PTMs have been able to do so only within short peptides.

In their paper, the Oxford scientists described their approach: We use electro-osmosis in an engineered charge-selective nanopore for the non-enzymatic capture, unfolding, and translocation of individual polypeptides of more than 1,200 residues. Unlabeled thioredoxin polyproteins undergo transport through the nanopore, with directional co-translocational unfolding occurring unit by unit from either the C or N terminus. Chaotropic reagents at non-denaturing concentrations accelerate the analysis.

PODCAST: In the Pacific’s depths, they found animals thought extinct since before the dinosaurs

The animals were thought to have gone extinct before the dinosaurs arrived on the scene.

Brian Kennedy, one of the lead scientists of NOAA’s deep sea expedition, said the stalked crinoid “are these big beautiful flower looking animals, but they grow on this long stalk. They’re called sea lilies, and that’s really what they look like a lot of times.”

“These were thought extinct, we thought they were a relic of pre-dinosaur times, and now we find they are one of the most common organisms in the deep sea.”

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