, Sodium chloride (NaCl), p.31434
, Magnesium chloride hexahydrate (MgCl2·6H2O), p.13152
, Polyethylene glycol (PEG), vol.600, p.87333
, catalog number: M8250)
, Magnesium sulfate (MgSO4)
, , p.131339
, Polyethylene glycol (PEG), vol.4000, p.95904
, Lithium sulfate (Li2SO4), p.203653
, Beryllium chloride (BeCl2) (Sigma-Aldrich, catalog number: 201197) Note: This product has been discontinued
, Sodium fluoride (NaF), p.71519
, Polyethylene glycol (PEG), vol.3350, p.202444
, Tri-potassium citrate, p.1722
, Yeast extract (Sigma-Aldrich, catalog number: Y1625)
, Tryptone plus, p.61044
, , p.9799
, EDTA-free protease inhibitor cocktail tablets (Roche Diagnostics, p.11873580001
, , p.104716
, Dithiothreitol (Soltec Ventures, catalog number: M112)
, Hydrochloric acid solution, pp.13-1683
, Immobilized Metal Affinity Chromatography (IMAC) binding and washing buffer
, IMAC elution buffer (see Recipes)
, Dialysis buffer (see Recipes)
, Size Exclusion Chromatography buffer for the phosphatase complex (SEC-P buffer)
, Size Exclusion Chromatography buffer for the phosphotransferase complex (SEC-PT buffer) Equipment
, Pipetman P2 single channel pipette (Gilson, catalog number: F144801)
, Pipetman P20 single channel pipette (Gilson, catalog number: F123600)
, Pipetman P200 single channel pipette (Gilson, catalog number: F123601)
, Pipetman P1000 single channel pipette (Gilson, catalog number: F123602) 5. 250 ml Erlenmeyer flasks, p.4110207
,
, L Erlenmeyer flasks (Marienfeld-Superior, p.4110217
, HisTrap immobilized metal affinity chromatography (IMAC) 5 ml column with Zn-NTA resin (GE Healthcare, pp.17-5248
, Superdex S75 26/600 (GE Healthcare, p.28989334
, Stirring magnet
, , pp.4053-8223
, Digital Ultrasonics Sonifier S-450 cell disruptor/homogenizer
, Multitron Standard incubation shaker (Infors HT)
, Minispin centrifuge (Eppendorf, model: MiniSpin ® , catalog number: 5452000018)
, Eppendorf, model: 5424 R, catalog number, pp.36-102
, Eppendorf, model: 5810 R, catalog number: 5811000010)
, Thermo Scientific TM , model: Sorvall Lynx4000, p.75006580
, Minipuls 3 peristaltic pump (Gilson, catalog number: F117604)
, , p.18664501
, EPS-301 power supply (GE Healthcare, model: EPS 301, catalog number
, Cary 50 Bio UV-visible spectrophotometer
, Alchemist DT)
, generator MicroMax-007HF (Rigaku, model: MicroMax-007 HF)
, Multilayer X-ray mirrors Varimax-HF
, Image plate area detector MAR345 ® (marXperts, model: mar345)
, SZX16 microscope, p.16
, , vol.330, p.330
, LG-PS2 light source, p.2
, Software 1. Produced diffraction datasets: a. Complex DesKC:DesR-REC in phosphotransferase state, low Mg 2+
, Complex DesKC:DesR-REC in phosphotransferase state, high Mg, vol.2
, Complex DesKC:DesR-REC in phosphotransferase state, high Mg 2+ and BeF3
, Complex DesKC:DesR-REC in phosphatase state
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