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Dear Manfred, attached are the postcript files. Vincent On 01/06/2017 10:19, Manfred S. Weiss wrote: > 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, Manfred > > Am 01.06.2017 um 10:12 schrieb vincent Chaptal: >> 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: >>> 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 1 >>> >>> >>> Crystal 2 is so different it might well have a dimer in a different >>> orientation. >> Yes, it could very well be the case. But wouldn't there be a peak as >> well in the SRF? >>> >>> 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.. >>> But doesnt mass spec or some such technique suggest whether there is >>> a dimer or not? >> 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. >> >> All the best >> Vincent >>> >>> Eleanor >>> Self Rotation Functions are a) hard to interpret and b) often >>> misleading! >>> >>> >>> On 31 May 2017 at 14:18, vincent Chaptal <[log in to unmask] >>> <mailto:[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 90.0000 >>> >>> 1 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 >>> >>> +33 4 37 65 29 01 <tel:+33%204%2037%2065%2029%2001> >>> >>> http://www.ibcp.fr >>> >> -- >> >> Vincent Chaptal, PhD >> >> Institut de Biologie et Chimie des Protéines >> >> Drug Resistance and Membrane Proteins Laboratory >> >> 7 passage du Vercors >> >> 69007 LYON >> >> FRANCE >> >> +33 4 37 65 29 01 >> >> http://www.ibcp.fr >> > -- > 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 -- Vincent Chaptal, PhD Institut de Biologie et Chimie des Protéines Drug Resistance and Membrane Proteins Laboratory 7 passage du Vercors 69007 LYON FRANCE +33 4 37 65 29 01 http://www.ibcp.fr