Alexey Nesvizhskii
@nesvilab.bsky.social
1.2K followers 170 following 43 posts
Godfrey D. Stobbe Professor of Bioinformatics at U of Michigan. Trained as a theoretical physicist, now focusing on proteomics and proteogenomics. https://fragpipe.nesvilab.org/
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nesvilab.bsky.social
Exciting news: We have released #FragPipe 23, and it's one of our biggest updates ever. Windows installer, support for TMT on Astral and timsTOF, TMT35, PTM site reports for DIA, improved Astral data handling in #MSFragger, improved diaTracer for diaPASEF data, better Skyline integration, and more!
Reposted by Alexey Nesvizhskii
maccoss.bsky.social
Proteomics Webinar: DIA with FragPipe, DIA-NN, and Skyline
Presenters: Eduard Sabidó and Brendan MacLean
When: Tuesday, September 16, 8am (Pacific Time)
Register Now ... skyline.ms/project/home...

#massspec #proteomics
Start Page: /home/software/Skyline/events/2025 Webinars/Webinar 26
skyline.ms
nesvilab.bsky.social
exact quote is "this site is being blocked due to DOJ Bulk Sensitive Data Regulations as the site is hosted in a Geo-Fenced country". I think replication of the data would be the only option for us in the foreseeable future.
nesvilab.bsky.social
The University of Michigan now blocks the iProX database, which is a part of the PRIDE consortium of mass spec data repositories. All requests to unblock were denied. Any other US universities in a similar situation? There is a lot of valuable MS proteomics data there no longer accessible to us.
Reposted by Alexey Nesvizhskii
fcyucn.bsky.social
It was a great pleasure to teach #FragPipe at the Biological Proteomics for Beginners workshop at #UCSD, sponsored by Thermo Fisher Scientific. We had a fantastic group of grad students, postdocs, and professors. Yes, I even got to teach UCSD professors how to analyze bottom-up proteomics data 😁
Reposted by Alexey Nesvizhskii
kusterlab.bsky.social
New preprint: We isolate peptide–RNA photo-crosslinks with tunable RNA chains from living cells for mass spec. This maps over 4,700 crosslinking sites across 744 proteins and offers the first glimpse into the RNA sequences in crosslinks by MS. Read here: doi.org/10.1101/2025...
Peptide-RNA photo-crosslinks with tunable RNA chain map protein-RNA interfaces
Photo-crosslinking mass spectrometry enables the identification of protein-RNA interactions in living cells, pinpointing interaction interfaces at single-amino acid resolution. However, current isolat...
doi.org
nesvilab.bsky.social
Dan Polasky is indeed a perfect teammate, and not only in our lab but also apparently as a … player in Kubb. I also want to use this opportunity to publicly congratulate Dan for being promoted to Research Assistant Professor starting September 1st!
gundrylab.bsky.social
“Champions” of Kubb at Dagstuhl! Dan Polasky is a great coder AND master gamer - perfect teammate! @nesvilab.bsky.social
nesvilab.bsky.social
I am not going to answer for “the field” but - in our tools - the short answer is yes. From introducing localization-aware open search, to Crystal-C artifact removal pubs.acs.org/doi/10.1021/..., to PTM-Shepherd summarization and visualization tools, we provide guardrails if not guidelines.
Crystal-C: A Computational Tool for Refinement of Open Search Results
Shotgun proteomics using liquid chromatography coupled to mass spectrometry (LC-MS) is commonly used to identify peptides containing post-translational modifications. With the emergence of fast databa...
pubs.acs.org
nesvilab.bsky.social
#MSFragger Open Search has been around for a while now and used by mass spec folks to screen for chemical artifacts and adducts, e.g. in chemoproteomics data. Happy to see it got 'discovered' by a broader community who are now reporting all sort of cool biological PTMs www.nature.com/articles/s41...
Nucleoside diphosphate kinase A (NME1) catalyses its own oligophosphorylation - Nature Chemistry
Our understanding of how post-translational modification—protein phosphorylation—impacts the complexity of eukaryotic signalling pathways is continuously expanding. Now, protein oligophosphorylation h...
www.nature.com
Reposted by Alexey Nesvizhskii
alisonchaves.bsky.social
It's now properly published. If you want to easily check important characteristics of your data before diving into complicated statistics, check out PSManalyst.

PSManalyst: A Dashboard for Visual Quality Control of FragPipe Results | Journal of Proteome Research pubs.acs.org/doi/10.1021/...
PSManalyst: A Dashboard for Visual Quality Control of FragPipe Results
FragPipe is recognized as one of the fastest computational platforms in proteomics, making it a practical solution for the rapid quality control of high-throughput sample analyses. Starting with version 23.0, FragPipe introduced the “Generate Summary Report” feature, offering .pdf reports with essential quality control metrics to address the challenge of intuitively assessing large-scale proteomics data. While traditional spreadsheet formats (e.g., tsv files) are accessible, the complexity of the data often limits user-friendly interpretation. To further enhance accessibility, PSManalyst, a Shiny-based R application, was developed to process FragPipe output files (psm.tsv, protein.tsv, and combined_protein.tsv) and provide interactive, code-free data visualization. Users can filter peptide-spectrum matches (PSMs) by quality scores, visualize protease cleavage fingerprints as heatmaps and SeqLogos, and access a range of quality control metrics and representations such as peptide length distributions, ion densities, mass errors, and wordclouds for overrepresented peptides. The tool facilitates seamless switching between PSM and protein data visualization, offering insights into protein abundance discrepancies, samplewise similarity metrics, protein coverage, and contaminants evaluation. PSManalyst leverages several R libraries (lsa, vegan, ggfortify, ggseqlogo, wordcloud2, tidyverse, ggpointdensity, and plotly) and runs on Windows, MacOS, and Linux, requiring only a local R setup and an IDE. The app is available at (https://github.com/41ison/PSManalyst.
pubs.acs.org
nesvilab.bsky.social
Conventional proteomics searches struggle with many modifications and fully open searches may be difficult to interpret. We introduce a "detailed" mass offset search in #MSFragger boosting interpretability and localization especially in complex cases (e.g. FPOP data): www.biorxiv.org/content/10.1...
nesvilab.bsky.social
Have you compared FragPipe on i9 vs comparably priced AMD threadripper? I put a request to our IT to get me one but they do not support core imaging of AMD machines. I am still recommending a good Xeon or i9 to folks who want a standard desktop to run FragPipe on a < $5k machine but want to test AMD
Reposted by Alexey Nesvizhskii
nicholasmccurtin.bsky.social
Don't tell PD, but I have quietly switched all of my analyses to Fragpipe. What an extremely powerful software. I'm often amazed at the sheer quantity of information I can get using Fragpipe. Thanks to everyone who recommended it.
nicholasmccurtin.bsky.social
Extremely niche troubleshooting request, but is anyone familiar with implementing MSFragger in Proteome Discoverer? A few of us keep seeing these error messages, but none of us are tech savvy enough to decode it. Every time we "solve" it, it happens again.. #proteomics #teammassspec
System.IO.FileNotFoundException: Could not find file 'D:\nick (temporary local analysis folder)\HeLa Pulldown\HeLa Pulldown\temp1593\50nM_OspF_02.pepXML.
File name: 'D:\nick (temporary local analysis folder)\HeLa Pulldown\HeLa
Pulldown\temp1593\50nM_OspF_02.pepXML'
at System.IO._Error.WinlOError(Int32 errorCode, String maybeFullPath) at System.IO.FileStream.Init(String path, FileMode mode, FileAccess access, Int32 rights, Boolean useRights, FileShare share, Int32 bufferSize, FileOptions options, SECURITY_ATTRIBUTES secAttrs, String msgPath, Boolean FromProxy, Boolean useLongPath, Boolean checkHost) at System.IO.FileStream.ctor(String path, FileMode mode, FileAccess access, FileShare share, Int32 bufferSize)
at System.XmI.XmIDownloadManager.GetStream(Uri uri, Credentials credentials, IWebProxy proxy, RequestCachePolicy cachePolicy) at System.Xml.Xm|Ur|Resolver.GetEntityUri absoluteUri, String role,
Type ofObject ToReturn)
at System.Xml.Xm/TextReaderImpl.FinishInitUriString0
at System.Xml.XmIReaderSettings.CreateReader(String inputUri,
Xm|ParserContext inputContext)
at System.Xml.Xm|Reader.Create(String inputUri)
at MSFraggerPDv30.PepXMLReader.ReadPepXML(String PepXMLPath)|
nesvilab.bsky.social
I miss Twitter a bit. With its outrageous commercials and all those gorgeously looking “Lucy564” people who seemed to wanted to follow me in droves. Ok, not really. I do occasionally check Bluesky but I only see posts from a few (mostly same) people. Lack of critical mass?
nesvilab.bsky.social
Wow, a record breaking number of the Nesvizhskii lab members attending #ASMS2025! 9 posters, 3 evening workshops, and one Bioinformatics Hub on #FragPipe. Plus multiple collaborative posters with other groups. See you in Baltimore! PS. Below is our recent group photo, including all those attending
nesvilab.bsky.social
The power of #MSFragger open search! “we applied the mass-tolerant search engine MSfragger and found that phosphorylation as well as ubiquitination were well preserved after XDNAX. To our great interest, we found an additional modification of 321 Da occurring only in the irradiated SILAC channel”
kusterlab.bsky.social
🚨Our new paper is online🚨
We use zero-distance⚡photo-crosslinking⚡to reveal direct protein-DNA interactions in living cells, enabling quantitative analysis of the DNA-interacting proteome on a timescale of minutes. #DNA #Chromatin #Proteomics
www.cell.com/cell/fulltex...
The human proteome with direct physical access to DNA
Zero-distance photo-crosslinking reveals direct protein-DNA interactions in living cells, enabling quantitative analysis of the DNA-interacting proteome on a timescale of minutes with single-amino-aci...
www.cell.com
Reposted by Alexey Nesvizhskii
embo.org
EMBO @embo.org · May 5
Don't miss out! Applications still open for EMBO Practical Course on Targeted proteomics: Advanced tools for biomedical research in Barcelona, Spain, 5 – 10 October 2025

Abstract submission & registration deadline: 15 May

meetings.embo.org/event/25-tar...
#EMBOtargetedProteomics #EMBOevents 🧪
Targeted proteomics: Advanced tools for biomedical research
Targeted proteomics technologies, and specially data-independent acquisition techniques, have revolutionized the landscape of proteomic research in the last decade offering researchers unprecedented …
meetings.embo.org
nesvilab.bsky.social
If there any #Sciex decision makers here on Bluesky - I urge you to reconsider. Skyline/Proteowizard support is not only important for your customers using these tools, but it also benefits other bioinformatics efforts that depend on these tools.
cashwood.proteaglyco.com
Disappointing to see, across both parties.
Sciex is no longer providing funding for Skyline/Proteowizard as part of the multi-vendor agreement to keep Skyline supported across all major vendors.
I can see the arguments from both sides, just a shame it's resulted in this.
Transcribed text:
Skyline Targeted Mass Spec Environment

Dear Skyline Users,


I would like to thank Agilent, Bruker, Shimadzu, Thermo Fisher, and Waters for their continued funding support for the Skyline project this year.  This industry/government/university partnership allows all of us to maximize the value of our efforts to provide better mass spectrometry tools to the field. In the current federal funding environment, industry support is critical to maintain our work.


Some instrument vendors, such as SCIEX, have not invested in the Skyline project this year. As a public university, we are limited in how we can support the private sector under state law.  We paused our work in Skyline and ProteoWizard on support for SCIEX instruments to remain in compliance with state law. While we hoped SCIEX would resume its funding support of the Skyline project for use with their instruments, and respect their decision not to provide funding, we cannot continue our work to support SCIEX in Skyline or ProteoWizard without SCIEX funding. We have informed the ProteoWizard Foundation of this development.


As always, we look forward to seeing many of you at ASMS or our Skyline course at UW this summer. We thank all of you for your continued support.

Brendan MacLean,

Skyline Project Lead
nesvilab.bsky.social
The tools remain free to academics under academic license. There has not been a discussion up to this point about changing that. Thanks to all our users, academic (now many thousands) and commercial, for their support!
nesvilab.bsky.social
Thank you so much for inviting us!
nesvilab.bsky.social
No, you still need to get a pre-release version from us. Email us please
Reposted by Alexey Nesvizhskii
dina-sch.bsky.social
Great presentation on #FragPipe and its capabilities. Definitely learned a lot!
Thank you @nesvilab.bsky.social for virtually stopping by to give a talk during our #proteomics users group meeting here @stanford.edu and for answering all our questions 😊

#massspec
#teammassspec
#StanfordProteomics