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On Jun 1, 2017, at 4:51 AM, vincent Chaptal <[log in to unmask]> wrote:Dear Manfred,
attached are the postcript files.
VincentOn 01/06/2017 10:19, Manfred S. Weiss wrote:[log in to unmask]" style="font-family: Helvetica; font-size: 14px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255);" class="">Dear Vincent,
it would be good if you post the postscript file as well.
It is called molrep_rf.ps or something like that.
Cheers, ManfredAm 01.06.2017 um 10:12 schrieb vincent Chaptal:[log in to unmask]" type="cite" class="">Thank you for your email.
the anisotropic resolutions of the datasets are 5.6-7.1A for the best and worst diffracting directions of the crystal without additive, and 4.0-5.8A for the crystal with additive.
The two crystals come from the same prep and same drop setup, only differ from the presence of the additive during crystallogenesis. They are indeed two different crystals, I would be curious to know more how to compare these two datasets together as I thought it was not possible with such different cell parameters.On 31/05/2017 17:12, Eleanor Dodson wrote:[log in to unmask]" class="">Yes, it could very well be the case. But wouldn't there be a peak as well in the SRF?Well - you dont give details o f resolution but the sovent content and the peak of 0.51 would suggest a possible dimer in crystal 1Crystal 2 is so different it might well have a dimer in a different orientation.[log in to unmask]" class="">I am not aware of a way to test by Mass Spec or other techniques the content of the ASU, I would be very interested if anyone can further my knowledge on this.But doesnt mass spec or some such technique suggest whether there is a dimer or not?I would try to see if there was a relationship between Xtal 1 and Xtal 2 - can tell you how I would do that if you are interested..
All the best
Vincent[log in to unmask]" class="">Self Rotation Functions are a) hard to interpret and b) often misleading!EleanorOn 31 May 2017 at 14:18, vincent Chaptal <[log in to unmask]> wrote:Dear all,
I need help interpreting results from a SRF; I am very naïve at interpreting them and would appreciate any pointer...
I have 2 crystals, before and after additive during crystallogenesis. They have different cell parameters, and I am wondering if I have a monomer or a dimer in the ASU, and if the additive changed this.
crystal w/o additive:
P21 114,5 107,6 134,6 beta=95,74
1monomer in the ASU = 80% solvent, 1 dimer in the ASU = 61% solvent. Note that this high solvent content agrees well with the fact that this is a membrane protein, and diffracts both to low resolution and very anisotropic...
SRF from Molrep:+------------------------------------------+ | theta phi chi P(i)/P(0)|+------------------------------------------+ | 1 0.00 0.00 0.00 1.00 || 2 148.48 0.00 180.00 0.51 || 3 161.29 0.00 180.00 0.31 || 4 105.34 180.00 180.00 0.30 || 5 74.78 -56.26 179.58 0.27 || 6 15.14 -163.78 180.00 0.25 || 7 67.61 -40.16 180.00 0.24 || 8 134.54 -180.00 180.00 0.19 || 9 72.33 -37.82 180.00 0.18 || 10 69.72 38.92 175.62 0.18 || 11 110.28 -141.08 175.62 0.18 || 12 96.58 101.90 179.79 0.18 || 13 67.04 -15.02 179.71 0.18 || 14 62.76 -15.52 179.45 0.17 || 15 117.26 164.48 179.45 0.17 || 16 68.39 -18.92 179.86 0.17 || 17 70.52 -16.51 180.00 0.17 || 18 24.05 -162.02 179.98 0.17 || 19 78.61 -36.36 180.00 0.17 || 20 83.17 78.42 173.34 0.16 || 21 96.83 -101.58 173.34 0.16 || 22 81.25 -75.03 179.73 0.16 || 23 81.81 -17.73 180.00 0.16 || 24 11.02 -142.22 180.00 0.16 || 25 76.14 -16.64 179.71 0.16 || 26 76.56 -16.93 180.00 0.16 || 27 162.29 32.44 180.00 0.16 || 28 10.03 -136.82 180.00 0.15 || 29 68.67 -58.10 179.49 0.15 || 30 111.33 121.90 179.49 0.15 || 31 62.99 -20.19 180.00 0.14 || 32 97.24 145.00 179.78 0.14 || 33 99.56 165.86 179.74 0.14 || 34 84.27 -82.68 180.00 0.14 || 35 84.51 72.76 174.42 0.13 |+------------------------------------------+ Crystal with additive: P21
CELL 117.2840 110.3330 155.6880 90.0000 93.4280 90.00001 monomer in the ASU = 84% solvent, 1 dimer = 68% solvent.SRF from Molrep:+------------------------------------------+ | theta phi chi P(i)/P(0)|+------------------------------------------+ | 1 0.00 0.00 0.00 1.00 || 2 59.55 0.00 180.00 0.36 || 3 156.11 0.00 180.00 0.29 || 4 10.00 -165.58 180.00 0.26 || 5 110.22 -180.00 180.00 0.26 || 6 12.97 -169.15 180.00 0.23 || 7 166.21 -9.06 180.00 0.22 || 8 81.64 -9.61 179.74 0.21 || 9 86.02 80.72 178.22 0.20 || 10 93.98 -99.28 178.22 0.20 || 11 5.96 -54.00 180.00 0.19 || 12 67.40 -10.81 180.00 0.19 || 13 87.10 -94.56 179.91 0.19 || 14 147.13 47.70 10.71 0.19 || 15 147.13 -47.70 10.71 0.19 || 16 87.45 91.04 155.74 0.19 || 17 92.55 -88.96 155.74 0.19 || 18 5.33 -63.00 180.00 0.19 || 19 104.60 169.61 179.59 0.19 || 20 88.01 -85.32 179.05 0.18 || 21 92.02 94.64 179.68 0.18 || 22 87.61 91.31 153.16 0.18 || 23 92.39 -88.69 153.16 0.18 || 24 6.65 -46.80 180.00 0.18 || 25 153.26 36.01 21.16 0.18 || 26 153.26 -36.01 21.16 0.18 || 27 63.11 -11.30 179.51 0.18 || 28 76.82 -5.09 179.61 0.18 || 29 87.69 91.37 148.72 0.18 || 30 92.31 -88.63 148.72 0.18 || 31 86.74 -9.44 179.90 0.17 || 32 85.83 80.82 148.49 0.17 || 33 94.17 -99.18 148.49 0.17 || 34 175.25 57.61 180.00 0.17 || 35 4.63 -90.00 180.00 0.17 |+-----------------------------Can I say that the peak at 0.51 in the crystal without additive is significant, concluding that there is probably a dimer in the ASU in that crystal, in contrast to the crystal with additive where no peak stands out, which would lean towards a monomer in the ASU? Thank you for your help. Best Vincent-------------+ --Vincent Chaptal, PhD
Institut de Biologie et Chimie des Protéines
Drug Resistance and Membrane Proteins Laboratory
7 passage du Vercors
69007 LYON
FRANCE
--Vincent Chaptal, PhDInstitut de Biologie et Chimie des ProtéinesDrug Resistance and Membrane Proteins Laboratory7 passage du Vercors69007 LYONFRANCE+33 4 37 65 29 01-- Dr. Manfred S. Weiss Macromolecular Crystallography Helmholtz-Zentrum Berlin Albert-Einstein-Str. 15 D-12489 Berlin Germany
Helmholtz-Zentrum Berlin für Materialien und Energie GmbH
Mitglied der Hermann von Helmholtz-Gemeinschaft Deutscher Forschungszentren e.V.
Aufsichtsrat: Vorsitzender Dr. Karl Eugen Huthmacher, stv. Vorsitzende Dr. Jutta Koch-Unterseher
Geschäftsführung: Prof. Dr. Bernd Rech (kommissarisch), Thomas Frederking
Sitz Berlin, AG Charlottenburg, 89 HRB 5583
Postadresse:
Hahn-Meitner-Platz 1
D-14109 Berlin
http://www.helmholtz-berlin.de--<crystal-with-additive_rf.pdf><crystal-without-additive_rf.pdf>Vincent Chaptal, PhDInstitut de Biologie et Chimie des ProtéinesDrug Resistance and Membrane Proteins Laboratory7 passage du Vercors69007 LYONFRANCE+33 4 37 65 29 01