Diethyl pyrocarbonate

Diethyl pyrocarbonate (DEP) [1609-47-8] M 162.1, b 38-40°/12mm, 160-163 /atm, d 1.119, Op 1.398. Dissolve in Et20, wash with dilute HCl, H2O, dry over Na2S04. filter, evaporate and distil the residue first in vacuo then at atmospheric pressure. It is soluble in alcohols, esters, ketones and hydrocarbon solvents. A 50% w/w soln is usually prepared for general use. Treat with great CAUTION as DEP irritates the eyes, mucous membranes and skin. [Boehm and Mehta Chem Ber 71 1797 1938 Thoma and Rinke Justus Liebigs Ann Chem 624 30 1959.]... 

The chloro atom of 2-[4-(6-chloronicotinoyl)benzyloxy]-3-methyl-4//-pyrido[l,2-n]pyrimidin-4-one, its 6-methyl derivative and 2-[4-(6-chlo-ronicotinoyl)benzylthio]-3-methyl-4//-pyrido[l,2-n]pyrimidin-4-one was replaced by a 4-piperidinopiperidino and 4-phenylpiperazino group with 4-piperidinopiperidine and 4-phenylpiperazine (96EUP733633). The carboxyl group of 2-[4-(4-carboxybenzoyl)benzyloxy]-3-methyl-4//-pyrido[l,2-n]pyrimidin-4-one, prepared by hydrolysis of methyl ester in DMF with 1 N NaOH, was reacted first with diethyl pyrocarbonate in DMF at room temperature and then with 4-phenylpiperazine and 4-piperidinopiperidine to give the appropriate amide derivatives (96EUP733633). 

The A-( 1 -alkoxyalkyl)carbamates are more simply obtained by a one-step synthesis on heating of the imine and diethyl pyrocarbonate in ethanol 57 58. In the case of a secondary alkyl substituent toluene appears to be the preferred solvent59. 

Reaction of Imines with Diethyl Pyrocarbonate General Procedure59" ... 

A solution of 0.9 mmol of the imine and l.2mmol of diethyl pyrocarbonate in 1.5 mL of dry ethanol (distilled from magnesium ethoxide) is stirred under nitrogen at 60 C for 24 h. Then the solvent is removed in vacuo and the residue flash chromatographed (silica gel, ethyl acetate/hexane 1 5) to give the product as an oil. 

Diethyl pyrocarbonate (DEPC)-treated water. Prepare as a 0.1% (v/v) solution and incubate at 37° for 18 hrs. Autoclave for 15 min to destroy excess DEPC. 

From Sigma 3-aminoethylcarbazole (AEC) acrylamide/bis-acrylamide (30%) 37.5 1 amino acids alumina bentonite benzamidine bovine fiver tRNA bovine serum albumin (BSA) creatine phosphate (CP) diethyl pyrocarbonate (DEPC) dithiothreitol (DTT) Escherichia coli MRE600 tRNA pyrophosphatase (Ppase) Ca++ salt of folinic acid, (5-formyl THF) IIHPHS K salt of phospho-enol pyruvic acid, (PEP) creatine phospho kinase (CPK) protease inhibitor cocktail for fungal and yeast extracts phenylmethylsulfonyl fluoride (PMSF) spermidine trihydrochloride Tween 20. 

The process of modification is shown in Fig. 13. Lys-52 is first derivatized with 4-thiobutanal and then a catalytic imidazole is bonded through a disulfide bridge into the active site. This can now act as a general base/nucleophile in the hydrolysis of [42], as was verified first by the pH-rate profile and then by complete deactivation of the antibody by diethyl pyrocarbonate (an imidazole-specific inactivating reagent). 

Characterization of the modified plastocyanins was by Inductively Coupled Plasma Emission Spectroscopy to analyze for Ru and Cu (1 1 ratio), and by HNMR spectroscopy. In the HNMR characterization the C2H resonance of His59 at 8.2 ppm is seen to be lost due to paramagnetic line broadening effect of the attached Ru(III), Fig. 13 [50]. In a further test it is known that the His59 s of both native plastocyanins react with diethyl pyrocarbonate (DEPC) to give an JV-ethoxyhistidine derivative, (12), which absorbs strongly at 238 nm (e 2750M- cm-i), Fig. 14 [133]. 

Fig. 14. The increase in absorbance at 238 nm (Ae 2750 M cm ) on DEPC (diethyl pyrocarbonate) modification of A. variabilis plasto-cyanin. Spectra were recorded at 5 min intervals [50]...

Subsequent to DNA template linearization, the procedures for in vitro RNA transcription reaction are set up. It is essential to avoid RNase contamination by using gloves, sterile glassware, and water devoid of RNase activity [treated with 0.1% diethyl pyrocarbonate (DEPC) and autoclaved]. 

Diethyl pyrocarbonate (DEPC) (Fluka Chemical Corp., Ronkonkoma, NY). DEPC-treated water 800 pL DEPC/L of distilled water stir overnight or at least 1 h autoclave (see Note 3). 

DIETHYL PYROCARBONATE DEPHOSPHO-CoA KINASE Dephospho-CoA pyrophosphorylase, PANTETHEINE-RHOSRHATE ADENYLYL-TRANSFERASE DEPOLARIZATION ACTION ROTENTIAL HYPERROLARIZATION DEPOLYMERIZATION PROCESSIVITY... 

ELECTROSTRICTION DIETHYL PYROCARBONATE Diferric-tyrosyl radical cofactor, REDOX-ACTIVE AMINO ACIDS... 

RIBOFLAVINASE RIBOFLAVIN KINASE RIBOFLAVIN SYNTHASE RIBOFLAVINASE RIBOFLAVIN KINASE RIBOFLAVIN SYNTHASE RIBONUCLEASE (or RNase) PSEUDOROTATION DIETHYL PYROCARBONATE RIBONUCLEASE F RIBONUCLEASE II RIBONUCLEASE III Ribonuclease inactivation,... 

DIETHYL PYROCARBONATE RIBONUCLEASE IV RIBONUCLEASE P Ribonuclease, pancreatic,... 

Equipment PCR machine, scintillation counter, tabletop centrifuge, temperature-controlled water baths, equipment for horizontal and vertical electrophoresis, UV-illuminator, phosphor imager, automatic DNA sequencer, vacuum dot-blot manifold (Schleicher and Schuell). PCR 0.5 ml hot-start mbes, aerosol resistant pipette rips, autoclaved Eppendorf tubes (all from Fischer Scientific, Brightwaters, NY) and glassware, diethyl pyrocarbonate (DEPC, Sigma)-treated solutions. 

Water treated with diethyl pyrocarbonate (DEPC) 0 05% (w/v) overnight at room temperature, and then autoclaved to remove DEPC... 

Diethyl propanedioate, d317 1,1-Diethylpropargylamine, d333 Diethyl pyrocarbonate, d326 Diethyl suberate, d324... 

Evidence for the involvement of histidine residues in the active site has been obtained from a number of experiments. Treatment with the histidine-selective reagent diethyl pyrocarbonate causes complete loss of activity in prolyl 4-hydroxylase (EC 1.14.11.2) [215] and 2,-4-dichlorophenoxyacetate dioxygenase (TfdA) (EC 1.13.11 group) [211], Site-directed mutagenesis of some of the conserved His residues in prolyl 4-hydroxylase [216] and aspartyl P-hydroxylase (EC 1.14.11.16) [217] also results in the loss of enzymatic activity. More recently, direct spectroscopic evidence for His coordination has been obtained in ESEEM studies of Cu(II)-substituted TfdA [218],... 

DEPC diethyl pyrocarbonate DIFP diisopropyl fluorophosphate DTNB 5,5 -dithiobisnitrobenzoic acid pCMB p-chloromercuribenzoate PMSF phenyl-methylsulfonyl fluoride TLCK N-tosyl-L-lysine chloromethylketone. 

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