Nucleotide Analogs - Jena Bioscience

JBS Methylation Kit
Surface engineering of biological macromolecules provides a powerful
technique to deal with proteins which are reluctant to crystallize or which
yield poorly diffracting crystals only. The JBS Methylation Kit offers a
straightforward tool for the selective methylation of lysine residues.
The method does not require laborious cloning/expression/purifi cation
but chemically substitutes the protons of the amino group of lysine
residues with methyl groups. The result is a surface-engineered protein
within 24 hours, ready for crystallization.
Each JBScreen Methylation Kit contains all necessary reagents for
six methylation experiments. All components provided are ready to use,
so just follow the manual step-by-step. No background in chemistry
necessary.
JBScreen Single Stocks
Single stock solutions of the JBScreen components, i.e. buffers, salts
and precipitants, are ideal for the optimization of your crystallization
conditions. Using the same chemicals as utilized in the JBScreens
ensures higher reproducibility of your experiments. JBScreen Single
Stocks are ready for use: the concentration is adjusted and they are
sterile fi ltered.
Buffer Solutions
Cat. No. Amount Price (€)
HEPES Sodium Salt pH 7.5, 1 M CS-604 50 ml 30,--
Imidazole HCl pH 8.0, 1 M CS-605 50 ml 25,--
MES Sodium Salt pH 6.5, 1 M CS-603 50 ml 40,--
Sodium Acetate pH 4.6, 1 M CS-601 50 ml 15,--
Sodium Citrate pH 5.6, 1 M CS-602 50 ml 15,--
Tris HCl pH 8.5, 1 M CS-606 50 ml 15,--
Precipitants
Ball-and-Stick
representation of a
methylated lysine residue.
Cat. No. Amount Price (€)
MPD, 100% (v/v) CS-628 50 ml 15,--
2-Propanol, 100% (v/v) CS-630 50 ml 15,--
Glycerol (anhydrous), 100% (v/v) CS-629 50 ml 15,--
Polyethylene Glycol 400, 100% (w/v) CS-619 50 ml 15,--
Polyethylene Glycol 1000, 50% (w/v) CS-620 50 ml 15,--
Polyethylene Glycol 3350, 50% (w/v) CS-621 50 ml 15,--
Polyethylene Glycol 4000, 50% (w/v) CS-622 50 ml 15,--
Polyethylene Glycol 6000, 50% (w/v) CS-623 50 ml 15,--
Polyethylene Glycol 8000, 50% (w/v) CS-624 50 ml 15,--
Polyethylene Glycol 550 MME, 50% (w/v) CS-625 50 ml 15,--
Polyethylene Glycol 2000 MME, 50% (w/v) CS-626 50 ml 15,--
Polyethylene Glycol 5000 MME, 50% (w/v) CS-627 50 ml 15,--
References:
Kobayashi et al. (2003) Refi ned structure and functional implications of trehalose synthase from
Sulfolobus acidocaldarius. J. Applied Glycoscience 50:1.
Kurinov et al. (2003) High resolution X-ray structure of potent anti-HIV pokeweed antiviral protein-III.
Biochem. Pharmacol. 65:1709.
Kurinov et al. (2000) X-ray crystallographic analysis of pokeweed antiviral protein-II after reductive
methylation of lysine residues. Biochem. Biophys. Res. Comm. 275:549.
Kobayashi et al. (1999) Crystallization and improvement of crystal quality for X-ray diffraction
of maltooligosyl trehalose synthase by reductive methylation of lysine residues. Acta Cryst. D
55:931.
Rayment (1997) Reductive alkylation of lysine residues to alter crystallization properties of proteins.
Methods in Enzymology 276 (12):171.
Rypniewski et al. (1993) Structural consequences of reductive methylation of lysine residues in
hen egg white lysozyme: an X-ray analysis at 1.8-A resolution. Biochemistry 21:9851.
Rayment et al. (1993) Three-dimensional structure of myosin subfragment-1: A molecular motor.
Science 261:50.
Cat. No. Amount Price (€)
JBS Methylation Kit CS-510 1 Kit 150,--
Salts
Crystallization Optimization
Further recommended reading:
Derewenda (2004) Rational protein crystallization by mutational surface engineering. Structure
12:529.
Cat. No. Amount Price (€)
Ammonium Acetate, 8.0 M CS-607 50 ml 20,--
Ammonium Sulfate, 3.5 M CS-608 50 ml 15,--
Lithium Chloride, 10.0 M CS-609 50 ml 20,--
Lithium Sulfate, 2.0 M CS-610 50 ml 20,--
Magnesium Chloride, 2.0 M CS-611 50 ml 20,--
Magnesium Sulfate, 2.5 M CS-612 50 ml 15,--
Potassium Chloride, 4.0 M CS-613 50 ml 20,--
Sodium Acetate, 3.0 M CS-614 50 ml 15,--
Sodium Chloride, 5.0 M CS-615 50 ml 15,tri-Sodium
Citrate, 1.6 M CS-616 50 ml 15,--
Sodium Malonate, 3.4 M CS-617 50 ml 25,--
Zinc Acetate, 1.0 M CS-618 50 ml 20,--
Macromolecular Crystallography
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