Protein sequencing articles from across Nature Portfolio

Homologous protein search is one of the most commonly used methods for protein analysis. Here, authors propose PLMSearch, a search method that takes only sequences as input and can search millions of protein pairs in seconds while maintaining sensitivity comparable to SOTA structure search methods.

A copper(II)-functionalized Mycobacterium smegmatis porin A nanopore enables direct identification of all 20 proteinogenic amino acids, one unnatural amino acid and two post-translational modifications, and shows potential for peptide discrimination and sequencing.

The lack of effective methods for mirror-image (d-) protein sequencing hampers the development of mirror-image biology systems and related applications. Now, total chemical synthesis of mirror-image trypsin enables the sequencing of long d-peptides and d-proteins, which may facilitate applications of d-peptides and d-proteins as potential therapeutic and informational tools.

Streets, Yosef, Robey and colleagues use multiomics analysis to generate a comprehensive timeline of the CD4⁺ and CD8⁺ T cell lineage commitment and identify sequential waves of TCR signaling that first initiate CD4⁺ T cell lineage differentiation and then CD8⁺ T cells lineage specification.

Developments in nanopore-based peptide detection and sequencing show promise of a breakthrough.

By using fixed charges to engineer a strong electroosmotic flow, we achieve the unidirectional transport of natural polypeptides across nanopores. Our approach enables native proteins to be transported enzymatically and non-enzymatically in the absence of denaturant and electrophoretic tags, with potential applications for protein sequencing.

The nanopore community is stepping toward a new frontier of single-molecule protein sequencing. Here, we offer our opinions on the unique potential for this emerging technology, with a focus on single-cell proteomics, and some challenges that must be overcome to realize it.

Mammalian cells have about 30,000 times as many protein molecules as mRNA molecules, which has major implications in the development of proteomics technologies. We discuss strategies that have been helpful for counting billions of protein molecules by liquid chromatography–tandem mass spectrometry and suggest that these strategies can benefit single-molecule methods, especially in mitigating the challenges posed by the wide dynamic range of the proteome.

Sequencing of proteins is a technically difficult task that typically requires digestion into short peptides before detection and identification. We developed a digestion-free method to chemically unfold and ‘scan’ full-length proteins through a nanopore, producing electrical fingerprints unique to individual protein molecules that are useful in their identification.