Hey #SciSky – let's settle something. Are you team "Western blot", or are you team "western blot"? Let me know. 🤓
Nicholas McCurtin, PhD 🧫 🏳️🌈
@nicholasmccurtin.bsky.social
PhD in Chemistry. Tufts University alum (Scheck Lab). Scientific support, sales, and marketing at HUABIO. Pop culture enthusiast, amateur chef, dilettante aesthete... (they/them) Website: https://nicholasmccurtin.com/
Cooking up some cool stuff over at HUABIO. I know I just defended a few months ago, but I'm already nostalgic for RAB1A. One of my favorite proteins to work with! But maybe that's because the westerns always looked great... 😆
HUABIO Research Inc 🧪🔬@huabioresearch.bsky.social · 2mo agoHappy Friday #SciSky – how was your week? Anything fun happen in the lab? Staying cool in the heatwaves? 🥵 We had a good week in Boston. Here's some immunofluorescence of our new RAB1A antibody, which I was really excited to see debut in our catalog as I worked with this protein a lot in my PhD!
In other news, I defended my dissertation last month, and I am beginning a new role at HUABIO as a scientific support, sales, and marketing specialist. After countless western blots, I am very excited to be on the other side of the antibody sales!
Ever wonder how bacteria hijack eukaryotic biochemical machinery to cause disease on a cellular level? Well, we finally have an activity-based method to profile how bacterial phosphothreonine lyases do just that. Read about my recent work in this area here! #teammassspec #chemsky #scisky
Profiling Selectivity for the Shigella Virulence Factor OspF
Phosphothreonine lyases are a small family of bacterial virulence factors that are secreted into host cells during infection by some Gram-negative pathogens. Phosphothreonine lyases catalyze an irreversible phosphate β-elimination that converts phosphothreonine (pThr) to dehydrobutyrine (Dhb) on host proteins. Here, we explore the substrate profile for OspF, a phosphothreonine lyase secreted during Shigella flexneri infection, which has been previously reported to modify mitogen-activated protein kinases (MAPKs) to silence the host immune response. In this work, we use a combination of in vitro assays with synthetic phosphopeptides and bottom-up chemoproteomic profiling to uncover the OspF substrates. These studies not only show that OspF exhibits selectivity within the MAPK family but also reveal that OspF modifies a wide range of cellular targets in cellular lysates and during Shigella flexneri infection. Using a nucleophilic phosphine chemical probe, we identified and validated new OspF targets beyond the MAPK family, including Rab1A and casein kinase 2β. Together, these findings provide valuable insights about OspF substrates in Shigella flexneri infection and highlight the need for further studies to reveal its full role in the context of Shigella flexneri pathogenesis. Moreover, these data underscore the potential utility of OspF and other phospholyases as novel chemical and synthetic biology tools for site-specific protein editing.
pubs.acs.org
This year's Black History Month Theme is A century of BHM Commemorations! It's been 100 years since the first observance of Black History Week.
Is anyone a fan of TMT? Doing my first quantitative proteomics experiment soon, and I'm intimidated 😭 #chemsky #teammassspec #proteomics
Ok, thanks everyone. Right now I am trying open search. I have some instrument time next week, so I'll try an collision energy curve then too as I still have some sample from this experiment.
Nicholas McCurtin, PhD 🧫 🏳️🌈@nicholasmccurtin.bsky.social · 12mo agoDoes anyone have advice for how to handle large (>500 Da) synthetic mass adducts for proteomic analysis? I have evidence of this chemical modification in my samples via western blot, but I am struggling localizing it via Orbitrap. #teammassspec #proteomics #chemsky
9am on a Saturday morning and it's a long weekend for me (took Monday off because I'm seeing Kali Uchis on Sunday), but I feel drawn to looking at the status of my Fragpipe and my LCMS QTOF runs...
an older woman in a blue shirt is standing in the woods
Alt: an older woman in a blue shirt is standing in the woods
media.tenor.com
Does anyone have advice for how to handle large (>500 Da) synthetic mass adducts for proteomic analysis? I have evidence of this chemical modification in my samples via western blot, but I am struggling localizing it via Orbitrap. #teammassspec #proteomics #chemsky
#BiochemSky sound off below ⬇️ , what is your favorite biomolecule? 🧬 #SciSky
I saw a reddit thread today about the best molecular biology papers that people have read, which led me to this paper. Extremely eloquently written, but every time I think about cancer deeply I am floored by it's complexities. www.cell.com/fulltext/S00...
The Hallmarks of Cancer
After a quarter century of rapid advances, cancer research has generated a rich and complex body of knowledge, revealing cancer to be a disease involving dynamic changes in the genome. The foundation ...
cell.com
Hey #phosphorylation folks! I've played around with some commercially available titanium dioxide resin for peptide-level enrichment, but I want to look at some phosphoproteins via western blot. What's the most cost effective way for protein-level phospho-enrichment? #chemsky #scisky
Trying out Alphafold for the first time. Let's see what the hype is about.
an older woman in a blue shirt is standing in the woods
Alt: jamie lee curtis
media.tenor.com
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.
Nicholas McCurtin, PhD 🧫 🏳️🌈@nicholasmccurtin.bsky.social · last yr.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
After a very long time of feeling like I've been banging my head against the wall because my PhD project has felt like a barren fishing experiment, I finally feel like a have at least one hypothesis that I can directly test to learn about my enzymes of interest.
Thanks everyone for input! We tried a hard reset on the system, and it's seemingly restored function... 🤞🏻
Nicholas McCurtin, PhD 🧫 🏳️🌈@nicholasmccurtin.bsky.social · last yr.Extreme rookie question, but our lab just had a power outage and now our #Orbitrap Exploris 240 is not acquiring any data. Here's what our HeLa digest standard looks like before and after outage. Anyone know what might be going on? #teammassspec #proteomics #chemsky #scisky
Extreme rookie question, but our lab just had a power outage and now our #Orbitrap Exploris 240 is not acquiring any data. Here's what our HeLa digest standard looks like before and after outage. Anyone know what might be going on? #teammassspec #proteomics #chemsky #scisky
Campus-wide power outage? That's fine. We'll go out for ice cream instead!
Thanks to everyone for the input in this thread. I'm convinced that it was SDS contamination. I switched to S-trap, and it seems to have resolved the problem. They are also delightful to work with! #chemsky #proteomics #teammassspec
Nicholas McCurtin, PhD 🧫 🏳️🌈@nicholasmccurtin.bsky.social · last yr.It's happening again! These are the same sample, just different biological replicates. What could make my peptides elute like that? #chemsky #proteomics #teammassspec
Breaking News: A federal judge ordered the Trump administration to release Rumeysa Ozturk, a Tufts University student whose arrest by plainclothed immigration agents in Massachusetts led to a public outcry.
Judge Orders Release of Tufts Student Detained by ICE
nytimes.com
Has anyone worked with stimulate the c-Jun N-terminal kinase (JNK) in tissue culture? I've been working with EGF to stimulate ERK #signaling, but I'm hoping to expand to JNK... Let me know if anyone has some thoughts! #scisky #chemsky #MAPK
I wrote a blog reflecting on my journey through several rounds of unprecedented times as a chemistry PhD student. Read it here if that interests you! #scisky #chemsky #scicom proteome.substack.com/p/chemistry-...
Chemistry in the time of instability
This is not about how easy it is to break a bond: a reflection on the unique circumstances particular to the 2020-2021 cohort of chemistry PhD students.
open.substack.com
A chilling thought that I have nowadays when I contact someone outside of my safe little bubble – "is it safe to keep my pronouns in my email signature?" – unfortunate that we're back to a time like this (if we ever even really left), but I keep them because I can't turn my back on my identity.
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
Rümeysa Öztürk hasn’t been given her asthma medication and has had three asthma attacks, putting her life in danger and possibly violating disability civil rights. New from me with @hurwitz.bsky.social for @motherjones.com. www.motherjones.com/politics/202...
ICE is reportedly violating detained Tufts student’s right to medical care
Democrats say that Rümeysa Öztürk's inhaler has been withheld and she has had repeated asthma attacks.
motherjones.com
It's happening again! These are the same sample, just different biological replicates. What could make my peptides elute like that? #chemsky #proteomics #teammassspec
Nicholas McCurtin, PhD 🧫 🏳️🌈@nicholasmccurtin.bsky.social · last yr.Really unusual signal in some recent samples I injected on Orbitrap Exploris... Anyone ever seen something like this before? #teammassspec #chemsky #proteomics
Sometime in September, a very robust #HPLC purification method I've worked with since 2021 randomly stopped working. With some help from the new instrumentation specialist in my department, finally diagnosed the issue. My beautiful peak is back at the retention time where it's supposed to be! 🥳
Happy #transdayofvisibility! We've always been here, and we'll always be here. 🏳️⚧️ ⚧️
Really unusual signal in some recent samples I injected on Orbitrap Exploris... Anyone ever seen something like this before? #teammassspec #chemsky #proteomics