Dear all,
There is an open postdoctoral position in my lab. We are interested in the
structure and function of integral membrane enzymes that catalyze protein
lipidation (Science, 2018, 359, eaao6326; PNAS., 2022 Feb
15;119(7):e2022050119) and transporters of transition metal ions (J Biol
Chem.
I would say that in a small survey (~5-6) of bacterial enzymes from our lab,
AlphaFold2 predicted exactly the constructs that were discovered by limited
proteolysis and/or sequence alignment for currently unpublished structures.
It’s a useful tool but I’ve also had two cases where it
Hi Scott,
I've used AF2 order/disorder prediction (based up pLDDT score) to decided upon
construct boundaries. We turned a non-expressing construct into a reasonably
well expressing construct based on the AF2 prediction.
It's part of my construct design process now.
HTH,
Dave
--
Dr David C.
It may also be useful to look at homologues of the protein of interest, as
sometimes Alphafold shows very different domain arrangements despite close
sequence similarity.
John
On Mon, Apr 4, 2022 at 3:17 PM Andrew Lovering
wrote:
> Hi Scott
> We have obtained a structure of a flexible
Dear Scott,
Fabulous question. I think a lot of people are thinking about this quite deeply
right now but our experience with single acylation and charge ladder work
(unpublished) was that even a single amino acid charge change had a profound
impact on crystallization outcome in the 1,536
The PROSS (Protein Repair One-Stop Shop) at:
https://pross.weizmann.ac.il/step/pross-terms
was used to both express and be able to crystallize human acetylcholinesterase
in E. coli, which prior to using PROSS had been impossible.
See paper:
Goldenzweig, A., Goldsmith, M., Hill, S. E., Gertman,
Hi Scott
We have obtained a structure of a flexible clamshell like fold only after using
a disulphide mutant to lock the domains based on a Rosetta Fold model.
Interestingly, Alphafold put the very same residues further apart (probably a
relevant "open" pose)
Andy
Hello CCP4,
Has anyone successfully used the available ML/AI protein folding tools to guide
crystallization construct design? Maybe you had a protein or domain that was
resistant to crystallization efforts and the folding algorithms predicted some
loops or termini that were disordered? Then
The Institute of Cancer Research (ICR), London, is one of the world’s most
influential cancer research institutes, with an outstanding record of
achievement dating back more than 100 years. We provided the first convincing
evidence that DNA damage is the basic cause of cancer, laying the
Please look at IRRMC (proteindiffraction.org) meeting WEB page:
https://bioreproducibility.org/media/irrmc_conference/
If you are interested, please register to the meeting. Please note that
meeting is a part of the US National Committee for Crystallography
workshop series on
exploring
Hi everyone,
I have a bacteria that has a dormant phase and an active phase in its
biphasic life cycle. It secretes a specific stress response protein "rpoS."
I wish to show that the accumulation of this protein during the dormancy
lifecycle (so early dormancy to late dormancy) is at the level of
Hi there,
Somehow I've missed the original email :) Sorry!
There are options for expressing really toxic genes, some of which have
already been mentioned and others perhaps not:
1. tight regulation of expression (promoter, repressor, other regulatory
elements, or a combination thereof). Beyond
Hi Andy,
just to follow up on Christian suggestion, which is exactly the way to go.
In case you are using an already pET based vector, simply try BL21-AI
(https://www.thermofisher.com/order/catalog/product/C607003), which has the T7
RNA polymerase under arabinose promoter should do the trick.
Dear colleagues,
my group is looking to hire a motivated postdoc with experience in cell
biology, a keen interest in proteomics, and the ability to communicate with
structural biologists.
It would be fantastic if you could forward the ad below to suitable candidates
and/or circulate it within
*Extended registration deadline*
Dear all,
We are pleased to announce our 2-dayspractical workshop on */Approaches
for in cellulo structural biology with X-rays /*from *May **16^th **to
**17**^th **2022*in Hamburg, Germany.
The workshop will provide practical on-site training on the
Hi Andy,
have you tried another promotor? Arabinose is much tighter, just to be sure
that it is really not leaking.
Cheers
Christian
On Mon, Apr 4, 2022 at 10:20 AM Andrew Lovering
wrote:
> Dear Board,
>
>
>
> Perhaps off-topic, but in the wider scope it’s relevant to many on here.
>
>
>
> We
Isn’t that what we all say every year on the day after March finishes?
Harry
> On 1 Apr 2022, at 23:46, Paul Emsley wrote:
>
> That, for the record, is more or less what Ralf said 18 years ago.
> On 01/04/2022 23:38, Pavel Afonine wrote:
>> It's April 1st today, isn't it? -;)
>>
>>
>> On
Dear Board,
Perhaps off-topic, but in the wider scope it's relevant to many on here.
We have a gene that we are able to clone, and propagate in DH5a etc
non-expression cells (hence nucleotide sequence is non-toxic)
But, when we attempt to transfer to an expression strain we get no colonies
We
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