A Chemical Proteomics Method to Quantify Cysteine S-Acylation S-acylation, often referred to as S-palmitoylation, is a reversible and dynamic posttranslational modification that corresponds to the addition of a long-chain fatty acid to cysteine (Cys) residues. Established mass spectrometry-based chemoproteomics methods have improved our understanding of the S-acylation proteome, notably by identifying hundreds of S-acylated proteins, sometimes with the modified Cys. However, the precise quantification of S-acylation levels for each Cys within a single sample remains challenging at the proteome level. Quantification of S-acylation levels is critical to further our understanding of protein S-acylation in cellular function and its role in health and diseases. We report here the development of an S-acylation quantification workflow based on the sequential labeling of free Cys and S-acylated Cys with isotopic labeling reagents. The workflow was extensively optimized, notably by comparing the number of sites identified with two alkyne-tagged Cys-reactive isotopic probes and four azido-tagged biotin-based capture reagents. By integrating this enhanced workflow with high-field asymmetric waveform ion mobility spectrometry (FAIMS) on LC–MS/MS instruments for the separation of labeled peptides, over 17,000 unique Cys could be quantified in biological samples. Application of the S-acylation quantification workflow to cellular proteomes allowed for the quantification of S-acylation levels in a HeLa proteome. We also identified dynamic S-acylation changes in response to autophagy induction.

A Chemical Proteomics Method to Quantify Cysteine S-Acylation | ACS Chemical Biology pubs.acs.org/doi/abs/10.1...

Mass Spectrometry Technology Specialist, Międzynarodowy Instytut Biologii Molekularnej i Komórkowej w Warszawie, Warszawa Job offer Mass Spectrometry Technology Specialist, Międzynarodowy Instytut Biologii Molekularnej i Komórkowej w Warszawie, Warszawa

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A Technical Evaluation of Plasma Proteomics Technologies Plasma proteomic technologies are rapidly evolving and of critical importance to the field of biomedical research. Here we report a technical evaluation of six notable plasma proteomic technologies – ...

I'm planning on running an olink pilot study so I'll need to get up to speed in this. I see the Coons Lab did a comparison

www.biorxiv.org/content/10.1...

Hey #TeamMassSpec, with all the super duper ultra fast TOF based DIA methods out there, how are you managing the throughput & depth on your old tribrids, QEs and Exploris? Any TMT aficionados out here getting 30spd, for instance, or even 15 with >8k proteins?

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The lack of symmetry is really annoying. The port wing root broke which has resulted in the offset. 😭

Am I being silly, but I didn't think somalogic used antibodies. I thought they used protein binding DNA?

Also consider double digestion. I'm currently running where a specific C is being targeted. The only way we could detect it was digesting with Trypsin and AspN together

The underside is usually pretty boring; designed to disappear into the sky; maybe painted with sky grey etc and that's all you would see hung up.

I have started to try and give them away to people. If you are ever in Cambridge, UK I can hook you up 😉

Where do people put their #scalemodels when complete? Mine ends up in the attic, unseen as I have no space to display

I follow back people, though I'm struggling to keep up when requests. I'll get there eventually

And more competition can only help in driving down costs

I kind of used Bens blog as my 'bot' to be honest. Its curated and i really just don't have the time between work, two young kids etc to read as much as I would like

They were also offering a Lumos at deep discount as well

Hey #TeamMassSpecv of you only have one mass spec and you are running TMT, what is your preferred number of fractions (is you balancing depth of coverage with instrument time). Currently we are at 16 though curious as to what others do

Thanks. Looks great by the way 👏

I have heard it this, but never tried it. Do you dilute the putty?

They were from an older kit that I found at a modelling show.

It's not daft, it's asked the same question (I am not a FAIMS expert). Their argument was that choosing only CV is too selective and will not produce results representative of the whole sample. To ensure we are not filtering out certain peptides, we need at least 2 CV's.

I recently completed one with folded Wings...... Is is very fragile

Thanks for the link. Think it will be a really good topic for a lit review session

Linkedin is also connected with my employer (my manager, people I manage, directors ect). I'm definitely so not feel as open for discussion on it. I find it very hard on Twitter to find things in interested in lately so hoping here works.

We have an Exploris. Instrument time is too limited to run on multiple injections. I want to see if we can run with FAIMS for all our workflows as taking it on/off is inefficient, but the team have said DIA requires two CV voltages

We are running out with some PRM stuff and we still do TMT. Currently not using it for DIA as we use DIAnn for analysis. My experience had been mixed.

Anyone doing #proteomics have any experience of using Attobodies (alamarbio.com/technology/a...). Looks like an interesting platform (in the same vein as olink or somalogic

Hi #TeamMassSpec. What are you thoughts on running DIA with FAIMS and only using one compensation voltage? (So instead of running with for example 40 + 60, we only ran with 50)

Dear new glyco friends, here’s a starter pack we can work on, so we can bootstrap some great community on here.

go.bsky.app/QTL6EqT

First of two funded PhD positions in the lab - study norovirus replication with mass spectrometry, and molecular virology!

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We had one in Cambridge in October. It was really nice to meet Proteomics/mass spec people in and around Cambridge (though there were some London who also attended) My understanding is that they will be having more in the future