John Burke
@jburkevic.bsky.social
Professor at the University of Victoria. Structural biologist studying phosphoinositides, membrane signalling, and nanobodies using HDX-MS, x-ray crystallography and cryo EM
Thanks to @alextoker.bsky.social for the motivation to submit this to JBC @asbmbjournals.bsky.social , a great journal, with this being one of the most straightforward and useful manuscript review processes I have gone through in a while
November 5, 2025 at 10:55 PM
Thanks to @alextoker.bsky.social for the motivation to submit this to JBC @asbmbjournals.bsky.social , a great journal, with this being one of the most straightforward and useful manuscript review processes I have gone through in a while
The students working on this were exceptional. Sushant just defended his PhD, and is on the biotech job market with exceptional skills in HDX-MS, protein biophysics, and biochemical assays, with Alex defending in about 15 months looking for cryo EM postdocs. Keep an eye out for their applications!
November 5, 2025 at 10:52 PM
The students working on this were exceptional. Sushant just defended his PhD, and is on the biotech job market with exceptional skills in HDX-MS, protein biophysics, and biochemical assays, with Alex defending in about 15 months looking for cryo EM postdocs. Keep an eye out for their applications!
This was a fun effort involving the Cobb lab for nanobody identification, the Hansen lab for smTIRF, and the Balla lab for cellular assays. This nanobody will be a great way to target selectively TTC7B containing PI4K complexes. We are happy to share, with the sequence in the supplementary as well
November 5, 2025 at 10:41 PM
This was a fun effort involving the Cobb lab for nanobody identification, the Hansen lab for smTIRF, and the Balla lab for cellular assays. This nanobody will be a great way to target selectively TTC7B containing PI4K complexes. We are happy to share, with the sequence in the supplementary as well
Finally in collaboration with Scott Hansen's lab we showed this nanobody blocked EFR3 mediated membrane recruitment using his single molecule TIRF approach, and with Tamas Balla's lab showed this nanobody blocked EFR3 mediated PM recruitment in cells
November 5, 2025 at 10:39 PM
Finally in collaboration with Scott Hansen's lab we showed this nanobody blocked EFR3 mediated membrane recruitment using his single molecule TIRF approach, and with Tamas Balla's lab showed this nanobody blocked EFR3 mediated PM recruitment in cells
This nanobody clearly blocked efr3 recruitment in a dose dependent fashion
November 5, 2025 at 10:37 PM
This nanobody clearly blocked efr3 recruitment in a dose dependent fashion
We specifically wanted this TTC7B selective nanobody to block the ability to bind EFR3, as this is the critical factor in PI4KA plasma membrane recruitment. The cryo EM structure showed a clear steric block of EFR3 binding (based on Sushant and Alex's structure from Science Advances last year)
November 5, 2025 at 10:36 PM
We specifically wanted this TTC7B selective nanobody to block the ability to bind EFR3, as this is the critical factor in PI4KA plasma membrane recruitment. The cryo EM structure showed a clear steric block of EFR3 binding (based on Sushant and Alex's structure from Science Advances last year)
Sushant and Damilola in my group were running the BLI instrument nonstop to generate all of these curves. Alex was able to solve the cryo EM structure of this nanobody bound to PI4KA-TTC7B
November 5, 2025 at 10:35 PM
Sushant and Damilola in my group were running the BLI instrument nonstop to generate all of these curves. Alex was able to solve the cryo EM structure of this nanobody bound to PI4KA-TTC7B
This is important as diseases are caused by hyperactivation of PI4KA, but complete loss is toxic. Therefore we reasoned that we might be able to selectively target unique PI4K regulatory complexes. We developed a nanobody that was highly TTC7B selective
November 5, 2025 at 10:34 PM
This is important as diseases are caused by hyperactivation of PI4KA, but complete loss is toxic. Therefore we reasoned that we might be able to selectively target unique PI4K regulatory complexes. We developed a nanobody that was highly TTC7B selective
We fully characterised all possible regulatory complexes that can form between all 6 possible combinations of PI4KA regulatory complexes containing EFR3, TTC7 and FAM126 isoforoms
November 5, 2025 at 10:32 PM
We fully characterised all possible regulatory complexes that can form between all 6 possible combinations of PI4KA regulatory complexes containing EFR3, TTC7 and FAM126 isoforoms
Very cool paper, nice work!
October 15, 2025 at 2:24 PM
Very cool paper, nice work!
This was part of a special issue to celebrate 50 years since the landmark review from Bob Michell on phosphoinositide signalling. Looking forward to reading the full cohort of papers from this issue
September 10, 2025 at 10:09 PM
This was part of a special issue to celebrate 50 years since the landmark review from Bob Michell on phosphoinositide signalling. Looking forward to reading the full cohort of papers from this issue