I am actually a big fan of using KGlu solutions for protein purification and have found it to be compatible with both IEX and DNA binding (for replication proteins) at concentrations that NaCl does not support.
On Thu, Jan 12, 2017 at 2:14 AM, Jon R Sayers <j.r.say...@sheffield.ac.uk> wrote: > Following on I read somewhere a while back that potassium conc in E. coli > is estimated in the 30-300 mM range (http://book.bionumbers.org/ > what-are-the-concentrations-of-different-ions-in-cells/) . In other more > extremophiles it can be higher (Extremophiles > <https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4339784/#>. 2015; 19(2): > 315–325.) Maybe our default buffers should contain K+ and Glu at such > high conc- though not compatible for IEX of course it would appear that > such conditions are physiological at least for intracellular bacterial > proteins. > > > Prof. Jon R Sayers FRSB > Dept. of Infection, Immunity & Cardiovascular Disease > University of Sheffield Medical School > Beech Hill Rd > Sheffield S10 2RX > United Kingdom, > > Tel +44 (0)114 215 9552 <+44%20114%20215%209552> > Fax +44 (0) 114 271 3892 <+44%20114%20271%203892> > Email j.r.say...@shef.ac.uk > http://www.sheffield.ac.uk/iicd/profiles/sayers > > > On 12 Jan 2017, at 09:13, Reza Khayat <rkha...@ccny.cuny.edu> wrote: > > I don't think this is taught in Biochem101. You didn't miss it. The > cytoplasm is quite viscous, like jello. > > > > Reza Khayat, PhD > Assistant Professor > City College of New York > Department of Chemistry > New York, NY 10031 > > ________________________________________ > From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK <CCP4BB@jiscmail.ac.uk>] > on behalf of Tim Gruene [tim.gru...@psi.ch <tim.gru...@psi.ch>] > Sent: Thursday, January 12, 2017 3:55 AM > To: CCP4BB@JISCMAIL.AC.UK <CCP4BB@jiscmail.ac.uk> > Subject: Re: [ccp4bb] Completely Off-Topic > > Dear JPK, > > I was not aware of the absolute numbers, but maybe they are little > suprising: > when your tinned food contains 'yeast extract' it is equivalent to > monosodium > glutamate, which is commonly used as flavour enhancing agent. > > I am not a chemist to worry about it, but yeast seems to have a fullfilling > life with it. > > Best, > Tim > > On Thursday, January 12, 2017 12:45:03 AM CET Keller, Jacob wrote: > > Dear Crystallographers, > > Was anyone else aware that in E coli the intracellular glutamate > concentration is ~100 mM? Also other cell types (yeast, mammalian) are 10s > mM. Anything to say about this? I learned of this just recently, and have > been amazed about it for more than a week. Did I miss this in Biochem 101? > Does it matter? > > JPK > > ******************************************* > Jacob Pearson Keller, PhD > Research Scientist > HHMI Janelia Research Campus / Looger lab > Phone: (571)209-4000 x3159 <(571)%20209-4000> > Email: kell...@janelia.hhmi.org<mailto:kell...@janelia.hhmi.org > <kell...@janelia.hhmi.org>> > ******************************************* > > > -- > -- > Paul Scherrer Institut > Tim Gruene > - persoenlich - > OFLC/102 > CH-5232 Villigen PSI > phone: +41 (0)56 310 5297 <+41%2056%20310%2052%2097> > > GPG Key ID = A46BEE1A > > > > > > Best wishes, > > > Prof. Jon R Sayers FRSB > Dept. of Infection, Immunity & Cardiovascular Disease > University of Sheffield Medical School > Beech Hill Rd > Sheffield S10 2RX > United Kingdom, > > Tel +44 (0)114 215 9552 <+44%20114%20215%209552> > Fax +44 (0) 114 271 3892 <+44%20114%20271%203892> > Email j.r.say...@shef.ac.uk > http://www.sheffield.ac.uk/iicd/profiles/sayers > > > >
