Multi-Pass, Single-Molecule Nanopore Reading of Long Protein Strands

Explore groundbreaking advancements in protein sequencing with this video on multi-pass, single-molecule nanopore technology! Jeff Nivala, Ph.D., Keisuke Motone, Ph.D., and Daphne Kontogiorgos-Heintz discuss the development of a method to read intact, long protein strands using nanopore sensors, overcoming limitations of current technologies like Edman degradation and mass spectrometry. By leveraging ClpX unfoldase to pull proteins through a CsgG nanopore on the Oxford Nanopore Technologies MinION R9.4 flow cell, Nivala’s group achieved single-molecule sensitivity to amino acids, allowing for the detection of post-translational modifications such as phosphorylation. This innovative approach paves the way for accurate protein barcoding, identification of proteoforms, and deeper insights into biological processes and disease mechanisms. Watch now to learn how nanopore technology is revolutionizing proteomics!
Key Highlights:
• Full-length protein sequencing of designed proteins using nanopores.
• Multi-pass rereading of protein molecules for increased accuracy.
• Detection of single amino acid substitutions and post-translational modifications.
• Potential applications in proteoform identification and therapeutic development.

For more details, read the full article on Nature: https://doi.org/10.1038/s41586-024-07...

Code for analyses is available on Github: https://github.com/uwmisl/PASTOR-sequ...

Custom MinION MinKNOW run scripts can be obtained from Oxford Nanopore Technologies on request.
00:00 Motivation for this work and grand challenges in proteomics - Jeff Nivala, Ph.D.
04:26 Experimental design and use - Keisuke Motone, Ph.D.
07:00 Analytical tools and results - Daphne Kontogiorgos-Heintz
09:34 Assessment of post-translational modifications and folded proteins - Keisuke Motone, Ph.D.
12:06 Closing remarks and looking toward de novo single-molecule protein sequencing using nanopores - Jeff Nivala, Ph.D.

#proteomics #nanopore #scienceandtechnology #sequencing #Biotechnology #PostTranslationalModifications