Dear all, Sorry for the confusion created, I did not mean that a protein would have fit in the small unit cell. My question was -
1. Why are there closely spaced spots arising in salt crystal? 2. If TCEP could crystallize in the condition, I have got a protein (same as this)+ligand (different ligand) complex in very close condition. (ligand size is within 500Da). Thank you Kavya On 2023-02-03 14:35, a.perra...@nki.nl wrote: > Hi Kavya, > > Try https://csb.wfu.edu/tools/vmcalc/vm.html > > This tells you that a 30kD protein simply does not fit the cell. > > I am pretty sure you crystallised the ligand, or TCEP actually. > > Also, if you look at the diffractions pattern, its clear the crystal > diffracts beyond 1.0A, diffraction spots are really very very very strong at > 2.0A. > >> On 3 Feb 2023, at 09:22, kavyashreem <kavyashr...@instem.res.in> wrote: >> >> Dear all, >> >> We crystallized a protein (30kDa) + ligand (by cocrystallization), in the >> condition 10%PEG3350, 50mM Zinc acetate. >> >> Protein was in the buffer 20mM HEPES, 150mM NaCl, 1uM ZnCl2, 4mM TCEP, pH 8. >> >> >> Crystal: Crystal: >> crystal under UV m >> >> <b06fc576.png> <e091c7fd.png> <8ef9453e.png> >> >> When we collected the data at an in-house facility, it looked something like >> this: >> >> <b903961d.png> >> >> The minimum resolution spot is around 9Ang and maximum ~2.2Ang. >> >> I have not come across a protein diffraction like this, nor of a salt. When >> I ran the gel for the incubated protein (protein+ligand), there was no >> degradation. >> >> Although, I was sure there is some problem with this image I tried >> processing, which could not be, But indexing showed a unit cell of 11Ang, >> 11Ang, 46Ang in P3. which was quite expected for two of the axes but not the >> third. >> >> Can anyone please shed some light on this diffraction image? >> >> How can it happen? >> >> Thank you >> >> Regards >> >> Kavya >> >> ------------------------- >> >> To unsubscribe from the CCP4BB list, click the following link: >> https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > > ------------------------- > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
