Co expression of binding partner perhaps ? And then tagging the other protein
partner ?
Jürgen
..
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street, W8708
For that purpose we’ve developed this expression system that may or may not be
useful to your specific problem:
http://www.ncbi.nlm.nih.gov/pubmed/23996492
J Mol Recognit.http://www.ncbi.nlm.nih.gov/pubmed/23996492# 2013
Oct;26(10):496-500. doi: 10.1002/jmr.2292.
Development of a multifunctional
Let me pick up Eleanor’s comment:
is there something like a crystallographer today ? I mean in the true sense ?
I think as a “crystallographer” you won’t be able to survive the next decade,
you need to diversify your toolset of techniques as pointed out in this article
, please do not discourage your colleagues, particularly very junior
ones, from posting to the CCP4BB. Some of the questions may appear quaint or
irrelevant but it is easy to simply ignore topics that are of no interest!
Eugene
On 13 February 2014 14:41, Bosch, Juergen
jubo
Hi Bingfa,
first suggestion would be to process your data with the anomalous flag turned
on - just in case you happen to have some metal bound coincidentally and you
happen to have collected at a decent wavelength to pickup some anomalous
scattering. ~30% of proteins in the PDB have a metal
If heat is a concern, you can place your Emulsiflex into a bin with lots of
ice. We have the version with cooling after lysis, but if you are paranoid you
can cool the whole thing.
It has the footprint of a 15” MacBook Pro but it’s way more expensive than that
:-)
Jürgen
[Advertisement on] http://www.janscientific.com/#!/gallery [Advertisement off]
Jürgen
..
Jürgen Bosch
Johns Hopkins University
Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street,
Emulsiflex C5
alternative freeze-thawing, the old way we used to do this spiking with some
Lysozyme
or grinding it in a LN2 cooled mortar, is very effective and pretty cost
effective, assuming you don’t pay for the LN2
Jürgen
..
Jürgen Bosch
Johns Hopkins University
Hey David,
the 1 nM Kd by SPR sounds fishy to me - did you do these measurements yourself ?
Unless this is an antibody-protein complex or nanobody-protein complex or
protein-small molecule complex, the value is too low (for a normal PPI, I would
expect 50 -500 nM). Possible explanation for the
Hi Wei,
make two objects of your ligand, display one as sphere and the other as stick.
Then change the transparency setting for the spheres.
Jürgen
On Jan 19, 2014, at 11:40 PM, Wei Shi
wei.shi...@gmail.commailto:wei.shi...@gmail.com wrote:
Hi all,
Please see attached Fig where they show the
Step1:
visit Protparam tool and CP your sequence, scroll down until you find the
extinction coefficient part. If OD280 is not close to 1 then make sure to take
that into account in your chromatogram say it is 0.7
step2: look at your mAU's
If your peak is 1000 mAU then you have 0.7 mg/ml in that
Didn't mlphare use to print those values in the log file ?
Jürgen
On Dec 15, 2013, at 4:29 PM, David Schuller
dj...@cornell.edumailto:dj...@cornell.edu wrote:
I have some SIRAS data of a known structure. I want to get the
isomorphous and anomalous occupancy and phasing power from my data.
Hi Acoot,
since they behave differently on IEX, they are different - you would introduce
heterogeneity into your crystallization setup, which usually is not a good idea
for crystallization in general.
Are you using Benzonucleases by any chance in your preparation and if not, that
may explain
2.8 is not a terrible resolution to try out Buccaneer or Parrot. Your terrible
R-factor might be due to a shift in residues perhaps ? Your After-building map
does not show much of a side chain density to judge if you are in frame or off.
But the elongated helix is in my eyes convincing enough.
another option might be the age of the screen perhaps ?
very silly suggestion/question: your pH meter is calibrated before you make
your measurements right ?
If you have a nano-pH probe (anything that can be added to a 96well reservoir
well) you could test what the final pH of your composition
What is the solution to this?
Hi Meisam,
you have it, it is just three molecules in the asu. Look at the overall crystal
lattice packing and see if you have contacts supporting each molecule. Generate
a large representation of your symmetry mates, I suspect you have a channel in
your crystal
yes, indeed.
visit this site and make sure you get all green lights
http://molprobity.biochem.duke.edu
Jürgen
On Oct 31, 2013, at 11:25 AM, Debasish Chattopadhyay wrote:
I was wondering if there is a way to generate a PDB validation report before
depositing the coordinates so that one can go
You do use Coot and look at the density plots right ?
Phenix will essentially do the same but in text form (unless you use the GUI).
You can also do this with Solve/Resolve and you can write your own script with
CCP4 available tools to do the same RSR fit diagram.
Jürgen
On Oct 31, 2013, at
We've had that leak as well some time ago. Dismantle and reassemble it is the
trick. Just a slight angle when tightening it will lead to that problem. This
is usually the case because of overtughtening the reservoir to the main part of
the machine. Keep it always straight when closing it again.
Protein-protein interaction?
protein-small molecule interactions ?
Epitope mapping of mABs ?
Could you specify what you would like to do, as different models are good for
different things.
Jürgen
On Oct 28, 2013, at 10:08 AM, Gang Dong wrote:
Dear all,
Could anyone tell me your experience with
BiaCore 3000 or if you can afford the T200.
Proteon XPR36 would be a cheaper Option and should work as well.
Jürgen
On Oct 28, 2013, at 10:54 AM, Gang Dong wrote:
We mainly measure protein-protein interactions (sometimes protein-small
molecules). Thanks! _Gang
From: Bosch, Juergen [mailto:jubo
check out the hidden .nx folder in your home directory.
You should have a folder named config.
You may have a .nxs file or .cfg file there just edit the file with a text
editor.
You probably need to change this value:
option key=Resolution value=1440x900 /
If you have a retina MacBook I guess
Have a look at these two papers:
http://www.ncbi.nlm.nih.gov/pubmed/17004709
http://www.ncbi.nlm.nih.gov/pubmed/19929835
When you say high DMSO, how much is that in % ? Do you know if your crystals
survive that much percentage DMSO even without ligand ?
Jürgen
..
Jürgen
Is your gene of interest smaller than 750bp ?
Then I would synthesize the gene with IDT and optimize it for E.coli that's 139$
Jürgen
..
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns Hopkins Malaria Research
But you have not tested in the beam if the visually happy crystals diffract
right ?
I would do this first before consuming your precious ligands in worthless drops.
Jürgen
On Sep 12, 2013, at 10:34 AM, Christopher Browning wrote:
Right now I've tried 5, 10 and 16% DMSO (due to variations in
What a statement !
Give reviewers maps, I agree however, what if the reviewer has no clue of these
things we call structures ? I think for those people table 1 might still
provide some justification. I would argue it should go into the supplement at
least.
Jürgen
Sent from my iPad
On Aug
Since we keep discussing resolution cutoffs and the benefits of not to include
all data etc.
I thought I would crowd source your opinion on this particular data set.
processed with XDS, here's the XSCALE.LP output:
SUBSET OF INTENSITY DATA WITH SIGNAL/NOISE = -3.0 AS FUNCTION OF RESOLUTION
Health
Department of Biochemistry Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street, W8708
Baltimore, MD 21205
Office: +1-410-614-4742
Lab: +1-410-614-4894
Fax: +1-410-955-2926
http://lupo.jhsph.edu
On Aug 28, 2013, at 11:32 AM, Bosch, Juergen wrote
Hi Jim,
all data is good data - the more data you have the better (that's what they say
anyhow)
Not everybody is adopting to the Karplus Diederich paper as quickly as you do.
And not to be confused with the Diederichs and Karplus paper :-)
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3689524/
Hi Mahesh,
if you use Refmac, then you can tell it to refine the twin fraction, no need to
tell it the twin law as Refmac will figure it out. If you use phenix, you
explicitly tell it the twin law and refine then with it. You can get the
possible twin laws by running phenix.xtriage and looking
increase your PEG3350 27% and keep the other components at their current
concentration. You can also add glycerol or ethylene glycol into the mix.
If you have multiple crystals then try various variants.
Jürgen
On Aug 23, 2013, at 1:52 PM, Uday Kumar wrote:
Hello
Can anyone suggest a
Yes, I'm also surprised why people run gradients for the capturing step ? How
often is your desired protein spread out throughout the whole gradient ? So
what's the point the of the gradient if in the end you merge the whole peak
again ?
Maybe for a pilot study to identify how much imidazole
, 2013-08-22 at 10:07 -0400, Bosch, Juergen wrote:
Yes, I'm also surprised why people run gradients for the capturing
step ?
Because we can. Joking aside, I've seen some examples where protein
eluted at relatively low imidazole and upon running the gradient there
remains some (minimal) overlap
mashed potatoes
On Aug 22, 2013, at 2:17 PM, Ed Pozharski wrote:
On Thu, 2013-08-22 at 13:58 -0400, Bosch, Juergen wrote:
well if we hit the timer after lysis, say via cell disruptor then I
have my eluted protein in less than 1 hour, including 40 minutes batch
binding.
then proceeds to wait six
How about low pH elution or EDTA as alternative ?
Jürgen
..
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street, W8708
Baltimore, MD 21205
Phone:
Dear George,
if #4 is correct, shouldn't he be able to get good SIRAS using the peak dataset
as HA and the last collected dataset as native
Jürgen
..
Jürgen Bosch
Johns Hopkins University
Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns
Well said Petri,
also how much PEG3350 do you have in your conditions ? More than 25% ? I'm
going after cryo-conditions at this point, you might want to replace your
PEG3350 with smaller PEGs or a mixture of PEG400 and PEG3350.
Almost sounds as if no optimization of the original conditions was
Hi Mahesh,
TESTGEN START 1 END 120 ANGLE 0.5
moving the detector further back or using 2theta and collecting more frames.
When you started shooting at your image you should have checked for overlaps
ahead of time.
I assume this to about 1.8 Å - maybe it would have been wiser to collect a 2.5
Å
for #2)
I'd suggest get some of those Mitigen loops that are titled. I assume you have
hexagonal plates as crystals and you really want to shoot along the thin area
of the crystal down the sixfold. With normal loops it's an art to get that
crystal to sit upright in the loop but not impossible
tilted is what I meant at an angle of e.g. 30 or 60 degrees. Works fine with
most SSRL beamlines except of the 12-2 microfocus - but that might have been
fixed in the meantime.
Jürgen
On Aug 16, 2013, at 1:57 PM, Bosch, Juergen wrote:
for #2)
I'd suggest get some of those Mitigen loops
Hi Kevin et al.
Buccaneer IS a great program and I like it very much.
I have a testcase here for my X-ray class where a trimer is in the asymmetric
unit, however all copies are different (not in sequence, but in space - think
of Calmodulin for example).
There is no option to force Buccaneer
Hi Sheena,
simple quickfix to your problem:
a) Run CAD and limit the output resolution on that mtz file so you don't run
into the memory allocation error of pointless.
b) try phenix.xtriage also if it is not a CCP4 program
Jürgen
..
Jürgen Bosch
Johns Hopkins University
FKBP ?
http://www.pnas.org/content/97/13/7096.full.pdf
Jürgen
On Aug 8, 2013, at 8:03 PM, Shiva Bhowmik wrote:
Dear All,
I am looking for references and/or example of substrate or ligand induced
oligomerization of enzymes related to activation.
Any help in this regard would be greatly
VROCS, www.eyesopen.comhttp://www.eyesopen.com
Jürgen
On Aug 7, 2013, at 9:03 AM, Tobias Beck wrote:
Dear CCP4bb,
I would like to calculate the shape complementarity of several protein-ligand
complexes (crystal structures with ligand available). This involves a set of
different proteins and
Hi Appu,
nothing really to worry about if you process your data with XDS or d*trek using
3D profile fitting.
You still should be able to get something useful out of this data.
If you look at 90.png (I'm attaching a zoomed area of your image), you see your
crystal is either split or you had two
Try here:
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=8031loc=ec_rcs
download the sdf and use the sdf to pdb converter
Or I think phenix can read in sdf and convert them
Jürgen
On Jul 17, 2013, at 9:35 AM, Wei Feng wrote:
Dear all,
Thank you for your advices.
I had tried to use MPD
Special position ?
sulfate-size ?
Jürgen
On Jul 16, 2013, at 11:35 AM, Wei Feng wrote:
Dear all,
I found some redundant density in my structure beween two molecule. (see
picture 1 and 2)
But I am not sure which ligand will be.
Dose everyone see this ligand before? If so, can you tell the PDB
product of reaction.
Vaheh
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Bosch,
Juergen
Sent: Tuesday, July 16, 2013 11:53 AM
To: CCP4BB@JISCMAIL.AC.UKmailto:CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Dose anyone see this ligand
can you elute off the column with low pH ? Or EDTA if you don't want imidazole
around ?
Jürgen
On Jul 14, 2013, at 10:29 AM, Raji Edayathumangalam wrote:
Thanks everyone for your responses. I definitely plan to save the flowthrough
so we'll see what happens. My protein has a His tag and I did
Hi Fei,
I assume you have edited the XDS.INP file to include the initially found space
group and the first refined cell ?
If not, then that would be my first choice to do. If that then still fails with
the default parameters, then I would start fixing some values instead of
refining all of
Anybody out there who knows how many people are subscribed to the BB ?
Just curious. And no please don't reply all :-)
Jürgen
..
Jürgen Bosch
Johns Hopkins University
Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns Hopkins Malaria
Hi Eleanor,
C2 - this was XDS lingo or Bravais talk :-)
Jürgen
** LATTICE SYMMETRY IMPLICATED BY SPACE GROUP SYMMETRY **
BRAVAIS-POSSIBLE SPACE-GROUPS FOR PROTEIN CRYSTALS
TYPE [SPACE GROUP NUMBER,SYMBOL]
aP [1,P1]
mP [3,P2] [4,P2(1)]
mC,mI
The other obvious conclusion would be that dataset #3 is a different protein
perhaps ?
How about pointless for your third dataset ?
Jürgen
On Jun 10, 2013, at 2:57 PM, Wei Shi wrote:
Hi all,
I was trying to solve the structure of a protein in several different datasets
using xds and phenix. I
Hi Joern,
have a look at these two papers (and probably KK is reading this too and can
chime in with the right set of coordinates),
Korotkov et al. Secretins: dynamic channels for protein transport across
membranes. Trends Biochem Sci (2011) vol. 36 (8) pp. 433-43
Reichow et al. Structure of
Hi Yuri,
http://fpocket.sourceforge.net
http://sts-fw.bioengr.uic.edu/castp/calculation.php
Just to name 2
Jürgen
On Jun 5, 2013, at 7:12 PM, Yuri Pompeu wrote:
Dear BB,
I am sorry for posting off-topic but it is hard not to ask when you know you
can get a good answer ;-)
I need to
Hi Tobias,
this may or may not work.
calculate a map for both proteins, normalize them, put them on an identical
cell and subtract one map from another.
The program that can do this is called mapman *gosh* a non-CCP4 thing.
Good luck,
Jürgen
On Jun 4, 2013, at 10:18 AM, Tobias Beck wrote:
seems to be down sorry.
you should be able to use crossec from ccp4
Jürgen
On Jun 1, 2013, at 6:47 PM, Edward A. Berry wrote:
Is Ethan Merritt's anomalous scattering page at:
http://www.bmsc.washington.edu/scatter/
down or moved, or the firewall I'm behind is blocking it?
I want to check
Instead of making the crystal thicker, why don't you make the beamsize smaller
? Aka microfocus
If this is your cryo-condition then I would work on it a bit more or change the
drop ratio of protein:reservoir so that you can use a higher PEG3350
concentration in your reservoir. I would aim for
Hi Klaus,
- small molecular weight PEG's e.g. 200 instead of DMSO, has the advantage of
also helping to cryo protect
- Methanol (only for dispensing the compound into wells) then allow to
evaporate and simply add your cryo-protected crystals, the hope is that
sufficient of your ligand goes
I dare say indefinitely. Your crystal life expectancy drops rapidly though with
somebody not remembering to fill up the dewar. Transferring crystals to pucks
etc. is another problem. I had some crystals which I could not remember what
they were (label broke off the cane) and we simply reshoot
You can use it as an MR model in Molrep for example. Or are you asking where to
get the actual EM data from ? That would be here:
http://www.ebi.ac.uk/pdbe/emdb/
Jürgen
On May 18, 2013, at 8:32 PM, LISA wrote:
Hi All,
The EM structure of a complex was published at 8A. If we can collect
This looks to me like a good attempt for GraphEnt. Give it a shot and you might
be positively surprised.
Jürgen
Sent from my iPad
On May 13, 2013, at 21:26, Dale Tronrud det...@uoxray.uoregon.edu wrote:
Sometimes a floppy bit of a protein is even more floppy in a
particular crystal form.
Hi Faisal,
whatever makes your protein happy is good. What type of experiment were you
thinking of ? Protein-protein interaction or protein-small molecule interaction
? In the latter case add some Tween20 or Brij-35 into the mix. A decent
overview can be found under this webpage for what to
Hi Afshan,
You mention:
The protein prep is same which was using to get the Hits.
So how old is your prep ? Was it stored at 4C or in LN2? Do you know if your
protein is stable under the conditions you stored it ?
How long after purification did it take you to get those crystals ?
How old is
I thought imosflm recognizes automatically the 2theta offset ?
Jürgen
..
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street, W8708
Baltimore, MD 21205
There's a second side to that.
Reviewers who can't get enough data and request even more when you submit a
decent paper with 18 pages of supplement for example.
Jürgen
On Mar 29, 2013, at 7:32 AM, Toufic El Arnaout wrote:
http://www.plosone.org/article/info:doi%2F10.1371%2Fjournal.pone.0053374
Hi Tom,
some suggestions for you:
You should calculate a selfrotation function and see if you can learn something
from it.
You should definitely run Matthews Coefficient and see if you get a brilliant
idea there
And congratulations to your first crystal structure it looks really great, for
a
get the twin law and either refine with phenix.refine twin_law=-h,-k,l or
whatever it suggests, or just add into your Refmac script the line TWIN and it
will figure out the twin law for you.
You can also detwin data but then you might be throwing away a lot of data.
We've now had two cases
Hi George,
this is probably a very stupid suggestion and you likely have tried it, but
I'll suggest the obvious nevertheless.
What happens to your .nitc file when you rename it to .itc can you read it in
Origin then ?
Jürgen
On Mar 24, 2013, at 6:39 AM, George Kontopidis wrote:
Chris,
Try PDBSum
Jürgen
On Mar 23, 2013, at 9:34 AM, Faisal Tarique wrote:
Dear all
I am working on a thermostable protein and i have read that the stability to
high temperature is due to various ionic interactions among the amino acid
residues of the protein itself..I request you all to tell me
If I understand your question correctly, you have a couple of atoms which you
could align to get the rotation translation then you can use these values
with maprot (CCP4) or mama (USF) to actually superimpose maps.
Jürgen
On Mar 21, 2013, at 11:29 PM, Chen Zhao wrote:
Dear all,
Does anybody
Why not thermal denaturation in the presence of Sypro Orange and a realtime PCR
machine ?
Crowther et al. Use of thermal melt curves to assess the quality of enzyme
preparations. Anal Biochem (2010) vol. 399 (2) pp. 268-275
Or (shameless advertisement):
Hain et al. Structural characterization
Yep,
mostly you should stay away from Tris as this is the worst buffer system when
playing with temperature changes. Tris for example has a ∆pKa/10˚C -0.31
Good, N.E. (1986) Biochemistry 5, 467
Jürgen
P.S. @Matthew, was this what you meant by the Good buffers often not ? or
just a
Thank you James, you should write a History book about the modern x-ray times.
Or better make one of those movies you are famous for.
Jürgen
On Mar 16, 2013, at 10:46 AM, James Holton wrote:
The first report of shooting a protein crystal at a synchrotron (I
think) was in 1976:
Going back to the initial question.
I would recommend looking at AFITT
http://www.eyesopen.com/afitt
Works like a dream (in certain cases).
Jürgen
P.S. I wish I had some stocks from them but I don't
..
Jürgen Bosch
Johns Hopkins University
Bloomberg School of Public Health
Yep, I agree calculate the average B per structure and divide each B by this
value, then multiply it by any value that is reasonable so you can visualize
color differences :-)
Jürgen
On Mar 4, 2013, at 2:16 PM, Jacob Keller wrote:
You only entertain addition+subtraction--why not use
yes - but keep in mind your protein is in the context of the crystal lattice,
so flexible regions in solution are likely to be stabilized in the crystal
lattice. So if you color by B also look at the symmetry mates.
And you should also submit both structures to the TLSMD server and look at
I assume you use CM5 chips ?
I further assume you run at pH 7.5 perhaps ? What's the pI of your analyte ?
7.5 ? Do you get significant binding to your reference cell under the
conditions you are running ?
You might get rebinding to your negatively charged surface and the dissociation
you are
Well that looks pretty real then. You might have wrong concentrations in one or
the other experiment perhaps hence the difference.
Jürgen
On Feb 20, 2013, at 5:45 PM, xianchi dong wrote:
Thanks a lot for the kind reply.
I used CM5 chip at pH 7.4. I am not quite sure about the PI of my protein
I assume nobody of you is running an actual Osx server ? I mean the upgrade to
a full server version of the commonly distributed normal Osx releases ?
I have not done it yet but I do think many of the issues mentioned regarding
NFS/NIS could be addressed there. Regarding the missing macpro
Current Mac minis outperform my 2009 models with which I am still happy. So
dual core I guess would be sufficient no need for upgrade on graphics card.
Jürgen
..
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns
Any reason for the Mac Mini over the iMac
A zalman monitor ir can you hook up a second monitor in stereo mode to your
iMac ?
Jürgen
..
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns Hopkins Malaria Research
What is the best procedure to use for weak anomalous signal
That opens up the can of worms which I'm happy to jump into.
We've had very good success in the years 2003-2009 with shelx for finding sites
(sometimes more than 1 trials) then force feeding them to sharp for phase
improvement. We
Surprised Mercedes didn't sued him for that :-)
@Mr. Emsley I assume you soon will be Sir Emsley after that marvelous
painting.mentioned in Harry's email.
Jürgen
..
Jürgen Bosch
Johns Hopkins University
Bloomberg School of Public Health
Department of Biochemistry Molecular
coot: show symmetry molecules, then save symmetry molecule and you have your
tetramer most likely
Jürgen
On Jan 10, 2013, at 7:48 PM, james09 pruza wrote:
Hi,
Which program outputs the symmetry operator (rotation and translation)? I have
a dimer in the asymmetric unit and need to know the
bash or any other shell script using http://www.ccp4.ac.uk/html/coordconv.html
Probably moleman2 http://xray.bmc.uu.se/usf/moleman_man.html can handle your
request too.
Jürgen
On Jan 7, 2013, at 12:48 AM, Teri Arman wrote:
Fractional Coordiantes to Orthogonal Coordinates and Vice Versa
Hi, I
can't you feed solve with those positions or Hyss or the Sharp interface ?
It's been too long ago when I faced this issue but I remember force-feeding one
of the existing programs at the time with blank coordinates to figure it out -
or was it mlphare ?
Too long ago probably 10 years by now.
the months renamed alphabetically.
On 12/21/12 03:23, Tom Murray-Rust wrote:
Hi Juergen,
Your scheme as printed has two J's - so January and July are indistinguishable!
I would suggest the letters should instead be JFMAYULGSOND.
On 21 Dec 2012, at 01:52, Bosch, Juergen
jubo
You'll see the zinc also at the Semet peak edge already check out Ethan's edge
plot web server. If you used his-tag purification methods it would be wiser to
collect passed the edge of either Ni or Co so you can distinguish them from Zn
sites.
Jürgen
..
Jürgen Bosch
Johns
It probably had a functional Firewall under Linux that's why it survived :-)
Jürgen
P.S. note to myself, don't ever give George my laptop, as it might be
mistreated - just for test purposes :-)
On Nov 20, 2012, at 12:57 PM, George M. Sheldrick wrote:
It is dual bootable Linux/Windows
George
Hi Pavel,
does this also work for symmetry related atoms ?
Jürgen
..
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street, W8708
Baltimore, MD 21205
Phone:
Bill I think that's crap.
I had issues on a 2005 MacBook Pro with inflating battery and it was replaced
(after about 6 months). There were troubles with those batteries and impurities
but mine still had apple care at that time and the batteries were exchangeable.
I have not heard of the build
HI Tim,
you should know better. German is the most precise language, hence all those
old German *gosh* books (for the younger readers of this board, there was a
time before pdf and Nook readers) for organic chemistry etc. from the 19th
century and older (Beilstein, Angewandte ...). And why was
What if your mild urea condition unfolds your protein from a globular to a
stretched elongated form ? You probably have done CD already to verify that
this is not the case.
Jürgen
..
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry
Hi Matthias,
I have not read your attached MS however Figure 2 is very good. Congrats to the
first author for a very captive and engaging view of your protein.
Jürgen
On Nov 12, 2012, at 4:24 AM, Matthias Wilmanns wrote:
I am apologizing for sending an unintended email to CCP4 via CC.
Well originally we got these in cacodylate and not much else would diffract.
http://www.pdb.org/pdb/explore/materialsAndMethods.do?structureId=2EPH
But nowadays I have another conditions, which does not require cacodylate and
works well with Hepes.
Jürgen
On Nov 9, 2012, at 7:26 AM, Frank von
Hi Frank,
I like your letter with the random initiative :-)
And then the thread by citing a regulation, which they probably never have
heard of. I would additionally add the regulation numbers for the dewar. There
are two or three IATA numbers that should be mentioned to indicate its in line
@Cynthia,
On Nov 4, 2012, at 10:58 AM, Cynthia Kinsland wrote:
Since you're seeing the MBP band, it sounds as if you're getting some cleavage.
If there were no cleave you would only see the fusion and TEV bands on the gel.
I think that is a wrong assumption.
He did not specify if he sees the
calculate an anomalous map, you should see the Zn signal even if you collected
at the SeMet peak.
Jürgen
..
Jürgen Bosch
Johns Hopkins University
Bloomberg School of Public Health
Department of Biochemistry Molecular Biology
Johns Hopkins Malaria Research Institute
615 North
If you calculate an edgeplot via Ethan's server:
http://skuld.bmsc.washington.edu/scatter/AS_form.html
you'll see that Zn @1Å has about 3 anomalous electrons whereas Ca less than 1,
so assuming occupancy of 1 the stronger anomalous signal should give you a
hint, second looking at the refined
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